Pyrrolo[1,2-a]pyrazine spla2 inhibitor

ABSTRACT

wherein R 1  is —(L 1 )—R 6  wherein L 1  is a divalent linking group of 1 to 18 atoms or the like, and R 6  is a carbocyclic ring substituted by at least one non-interfering substituent or the like; R 2  is C1 to C3 alkyl, C3 to C4 cycloalkyl or the like group; R 3  is —(L 2 )-(acidic group); R 4  and R 5  are hydrogen atoms, non-interfering substituents, carbocyclic groups or the like; R A  is —C(═X)—C(═X)—NH 2  or the like; and X is independently oxygen atom or sulfur atom; the prodrugs thereof, their pharmaceutically acceptable salts, or their solvates, and a composition for inhibiting sPLA 2  containing them as effective ingredients.

TECHNICAL FIELD

The present invention relates to a pyrrolo[1,2-a]pyrazine derivativeeffective for inhibiting sPLA₂-mediated fatty acid release.

BACKGROUND ART

sPLA₂ (secretory phospholipase A₂) is an enzyme that hydrolyzes membranephospholipids and has been considered to be a rate-determining enzymethat governs the so-called arachidonate cascade where arachidonic acid,the hydrolysis product, is the starting material. Moreover,lysophospholipids that are produced as by-products in the hydrolysis ofphospholipids have been known as important mediators in cardiovasculardiseases. Accordingly, in order to normalize excess functions of thearachidonate cascade and the lysophospholipids, it is important todevelop compounds which inhibit the liberation of sPLA₂-mediated fattyacids (for example, arachidonic acid), namely, compounds which inhibitthe activity or production of sPLA₂. Such compounds are useful forgeneral treatment of symptoms, which are induced and/or sustained by anexcess formation of sPLA₂, such as septic shock, adult respiratorydistress syndrome, pancreatitis, injury, bronchial asthma, allergicrhinitis, chronic rheumatism, arteriosclerosis, cerebral apoplexy,cerebral infarction, inflammatory colitis, psoriasis, cardiacinsufficiency, cardiac infarction, and so on. The participation of sPLA₂is considered to be extremely wide and, besides, its action is potent.

There are known, as examples of sPLA₂ inhibitor, indole derivatives inEP-620214 (JP Laid-Open No. 010838/95), EP-620215 (JP Laid-Open No.025850/95), EP-675110 (JP Laid-Open No. 285933/95), WO 96/03376, and WO99/00360; indene derivatives in WO 96/03120; indolizine derivatives inWO 96/03383; naphthalene derivatives in WO 97/21664 and WO 97/21716;tricyclic derivatives in WO 98/18464; pyrazole derivatives in WO98/24437; phenylacetamide derivatives in WO 98/24756; phenyl glyoxamidederivatives in WO 98/24794; pyrrole derivatives in WO 98/25609.

DISCLOSURE OF INVENTION

The present invention provides pyrrolo[1,2-a]pyrazine derivatives havingsPLA₂ inhibiting activity and being useful for treatment of septicshock, adult respiratory distress syndrome, pancreatitis, injury,bronchial asthma, allergic rhinitis, chronic rheumatism, arterialsclerosis, cerebral hemorrhage, cerebral infarction, inflammatorycolitis, psoriasis, cardiac failure, and cardiac infarction.

The present invention relates to a compound represented by the formula(I):

wherein R¹ is hydrogen atom or a group selected from (a) C6 to C20alkyl, C6 to C20 alkenyl, C6 to C20 alkynyl, carbocyclic groups, andheterocyclic groups, (b) the groups represented by (a) each substitutedindependently with at least one group selected from non-interferingsubstituents, and (c) —(L¹)—R⁶ wherein L¹ is a divalent linking group of1 to 18 atom(s) selected from hydrogen atom(s), nitrogen atom(s), carbonatom(s), oxygen atom(s), and sulfur atom(s), and R⁶ is a group selectedfrom the groups (a) and (b);

R² is hydrogen atom, or a group containing 1 to 4 non-hydrogen atoms;

R³ is —(L²)—(acidic group) wherein L² is an acid linker having an acidlinker length of 1 to 5;

R⁴ and R⁵ are selected independently from hydrogen atom, non-interferingsubstituents, carbocyclic groups, carbocyclic groups substituted with anon-interfering substituent(s), heterocyclic groups, and heterocyclicgroups substituted by a non-interfering substituent(s); and

R^(A) is a group represented by the formula:

wherein L⁷ is a divalent linker group selected from a bond or a divalentgroup selected from —CH₂—, —O—, —S—, —NH—, or —CO—, R²⁷ and R²⁸ areindependently hydrogen atom, C1 to C3 alkyl or a halogen; X and Y areindependently an oxygen atom or a sulfur atom; and Z is —NH₂ or —NHNH₂;the prodrugs thereof; or their pharmaceutically acceptable salts; ortheir solvates.

Preferred subclass of compounds of formula (I) are those where for R¹ isthe divalent linking group —(L¹)— is a group represented by any one ofthe following formula (Ia) or (Ib) or (Ic):

wherein Q¹ is a bond or any one of the divalent groups (Ia) or (Ib) andeach R³⁶ is independently hydrogen atom, C1 to C8 alkyl, C1 to C8haloalkyl, or C1 to C8 alkyloxy. Particularly preferred as the linkinggroup —(L¹)— of R¹ is an alkylene chain of 1 or 2 carbon atoms, namely,—(CH₂)— or —(CH₂CH₂)—.

Preferred sPLA₂ inhibitor compounds of the invention are thoserepresented by the formula (II):

wherein R⁷ is hydrogen atom or —(CH₂)m—R¹² wherein m is an integer from1 to 6, and R¹² is (d) a group represented by the formula:

wherein a, c, e, n, q, and t are independently an integer from 0 to 2,R¹³ and R¹⁴ are independently selected from a halogen, C1 to C10 alkyl,C1 to C10 alkyloxy, C1 to C10 alkylthio, aryl, heteroaryl, and C1 to C10haloalkyl, α is an oxygen atom or a sulfur atom, L⁵ is —(CH₂)v—, —C═C—,—C≡C—, —O—, or —S—, v is an integer from 0 to 2, β is —CH₂— or —(CH₂)₂—,γ is an oxygen atom or a sulfur atom, b is an integer from 0 to 3, d isan integer from 0 to 4, f, p, and w are independently an integer from 0to 5, g is an integer from 0 to 2, r is an integer from 0 to 7, and u isan integer from 0 to 4, or is (e) a member of (d) substituted with atleast one substituent selected from the group consisting of C1 to C6alkyl, C1 to C6 alkyloxy, C1 to C6 haloalkyloxy, C1 to C6 haloalkyl,aryl, and a halogen;

R⁸ is C1 to C3 alkyl, C1 to C3 alkenyl, C3 to C4 cycloalkyl, C3 to C4cycloalkenyl, C1 to C2 haloalkyl, C1 to C3 alkyloxy, or C1 to C3alkylthio;

R⁹ is —(L³)—R¹⁵ wherein L³ is represented by the formula:

wherein M is —CH₂—, —O—, —N(R²⁴)—, or —S—, R¹⁶ and R¹⁷ are independentlyhydrogen atom, C1 to C10 alkyl, aryl, aralkyl, alkyloxy, haloalkyl,carboxy, or a halogen, and R²⁴ is hydrogen atom or C1 to C6 alkyl, andR¹⁵ is represented by the formula:

wherein R¹⁸ is hydrogen atom, a metal, or C1 to C10 alkyl, R¹⁹ isindependently hydrogen atom, or C1 to C10 alkyl, and t is an integerfrom 1 to 8;

R¹⁰ and R¹¹ are independently hydrogen atom or a non-interferingsubstituent selected from hydrogen, C1 to C8 alkyl, C2 to C8 alkenyl, C2to C8 alkenyl, C7 to C12 aralkyl, C7 to C12 alkaryl, C3 to C8cycloalkyl, C3 to C8 cycloalkenyl, phenyl, tolyl, xylyl, biphenyl, C1 toC8 alkyloxy, C2 to C8 alkenyloxy, C2 to C8 alkynyloxy, C2 to C12alkyloxyalkyl, C2 to C12 alkyloxyalkyloxy, C2 to C12 alkylcarbonyl, C2to C12 alkylcarbonylamino, C2 to C12 alkyloxyamino, C2 to C12alkyloxyaminocarbonyl, C1 to C12 alkylamino, C1 to C6 alkylthio, C2 toC12 alkylthiocarbonyl, C1 to C8 alkylsulfinyl, C1 to C8 alkylsulfonyl,C2 to C8 haloalkyloxy, C1 to C8 haloalkylsulfonyl, C2 to C8 haloalkyl,C1 to C8 hydroxyalkyl, —C(O)O(C1 to C8 alkyl), —(CH₂)z—O—(C1 to C8alkyl), benzyloxy, aryloxy, arylthio, —(CONHSO₂R²⁵), —CHO, amino,amidino, halogen, carbamyl, carboxyl, carbalkoxy, —(CH₂)z—CO₂H, cyano,cyanoguanidinyl, guanidino, hydrazide, hydrazino, hydrazido, hydroxy,hydroxyamino, iodo, nitro, phosphono, —SO₃H, thioacetal, thiocarbonyl,or carbonyl, R²⁵ is C1 to C6 alkyl or aryl, z is an integer from 1 to 8;and R^(B) is a group represented by the formula:

wherein Z is the same as defined above; the prodrugs thereof, or theirpharmaceutically acceptable salts, or their solvates.

When the above b, d, f, p, r, u, and/or w are 2 or more, a plural numberof R¹³ or R¹⁴ may be different from one another. When R¹³ is asubstituent on the naphthyl group, the substituent may be substituted atany arbitrary position on the naphthyl group.

The invention also relates to preferred compounds represented by formula(I) or (II) the prodrugs thereof, or their pharmaceutically acceptablesalts, or their solvates, wherein said R¹ and R⁷ are represented by theformula:

wherein R¹³, R¹⁴, b, d, f, g, p, r, u, w, α, β, and γ are the same asdefined above, L⁶ is a bond, —CH₂—, —C═C—, —C≡C—, —O—, or —S—.

When the above b, d, f, p, r, u, and/or w are 2 or more, a plural numberof R¹³ or R¹⁴ may be different from one another. When R¹³ is asubstituent on the naphthyl group, the substituent may be substituted atany arbitrary position on the naphthyl group.

The invention also relates to preferred compounds represented by formula(I) and (II), the prodrugs thereof, or their pharmaceutically acceptablesalts, or their solvates, wherein for the formula (I) and (II)respectively the substituent R² or R⁸ is selected from C1 to C3 alkyl orC3 to C4 cycloalkyl.

The invention also relates to a preferred compound of formula (I) or(II), the prodrugs thereof or their pharmaceutically acceptable salts,or their solvates, wherein the L² and L³ are —O—CH₂—.

The invention also relates to a preferred compound represented by theformula (III):

wherein R²⁰ is a group represented by the formula:

wherein L⁶, R¹³ R¹⁴, b, d, f, g, p, r, u, w, α, β and γ are the same asdefined above;

R²¹ is C1 to C3 alkyl or C3 to C4 cycloalkyl;

L⁴ is —O—CH₂—, —S—CH₂—, —N(R²⁴)—CH₂—, —CH₂—CH—, —O—CH(CH₃)—, or—O—CH((CH₂)₂Ph)— wherein R²⁴ is hydrogen atom or C1 to C6 alkyl and Phis phenyl;

R²² is —COOH, —SO₃H, or P(O)(OH)₂;

R²³ is hydrogen atom, C1 to C6 alkyl, C7 to C12 aralkyl, C1 to C6alkyloxy, C1 to C6 alkylthio, C1 to C6 hydroxyalkyl, C2 to C6haloalkyloxy, halogen, carboxy, C1 to C6 alkyloxycarbonyl, aryloxy,arylthio, a carbocyclic group, or a heterocyclic group; and R^(B) is thesame as defined above; the prodrugs thereof; or their pharmaceuticallyacceptable salts; or their solvates.

When the above b, d, f, p, r, u, and/or w are 2 or more, a plural numberof R¹³ or R¹⁴ may be different from one another. When R¹³ is asubstituent on the naphthyl group, the substituent may be substituted atany arbitrary position on the naphthyl group.

The invention also relates to most preferred compounds represented bythe formula (IV):

wherein R^(20,) R²¹, R^(23,) and R^(B) are the same as defined above;and k is an integer from 1 to 3; the prodrugs thereof; or theirpharmaceutically acceptable salts; or their solvates.

The invention also relates to a preferred compound, the prodrugsthereof, or their pharmaceutically acceptable salts, or their solvatesas described in formula (III) wherein L⁴ is —O—CH₂—.

The invention further relates to a preferred compound, the prodrugsthereof, or their pharmaceutically acceptable salts, or their solvatesas described in formula (I), (II), (III), or (IV), wherein R^(A) andR^(B) are —COCONH₂—.

The invention also relates to preferred compounds formula (I), (II),(III), or (IV), the prodrugs thereof, or their pharmaceuticallyacceptable salts, or their solvates wherein R^(A) and R^(B) are—CH₂CONH₂—.

The invention further relates to preferred compounds of formula (I),(II), (III), or (IV), the prodrugs thereof, or their pharmaceuticallyacceptable salts, or their solvates wherein R^(A) and R^(B) are—CH₂CONHNH₂—.

The invention also relates to preferred compounds of formula (I), (II),(III), or (IV) in the form of ester type prodrug.

The invention further relates to specific preferred sPLA₂ inhibitorcompounds of formula (I), (II), (III), or (IV), namely, apyrrolo[1,2-a]pyrazine compound selected from the group consisting of:

[6-Benzyl-7-ethyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyacetic acid,

[6-Cyclohexylmethyl-7-ethyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyacetic acid,

[7-Ethyl-6-(3-methoxybenzyl)-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyaceticacid,

[6-(Benzo[b]thiophen-6-ylmethyl)-7-ethyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyacetic acid,

[6-Benzyl-7-ethyl-3-methyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyaceticacid,

[7-Ethyl-6-(4-fluorobenzyl)-3-methyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyacetic acid,

[6-(2-Biphenylmethyl)-7-ethyl-3-methyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyacetic acid,

[6-Cyclopentylmethyl-7-ethyl-3-methyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyaceticacid,

[6-(2-Benzyl)benzyl-7-ethyl-3-methyl-8-oxamoylpyrrolo [1,2-a]pyrazin- 1-yl]oxyacetic acid,

[7-Ethyl-6-(2-(4-fluorophenyl)benzyl)-3-methyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyaceticacid,

[7-Ethyl-6-(3-fluorobenzyl)-3-methyl-8-oxamoylpyrrolo [1 ,2-a]pyrazin-1-yl]oxyacetic acid,

[6-Benzyl-7-ethyl-3-isopropyl-8-oxamoylpyrrolo [1,2-a]pyrazin-1-yl]oxyacetic acid,

[6-Benzyl-3,7-diethyl-8-oxamoylpyrrolo [1,2-a]pyrazin-1-yloxyaceticacid,

[6-Benzyl-7-ethyl-8-oxamoyl-3-phenylpyrrolo[1,2-a]pyrazin-1-yl]oxyaceticacid,

[6-Benzyl-7-ethyl-3-isobutyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyaceticacid,

[3,6-Dibenzyl-7-ethyl-8-oxamoylpyrrolo[1,2-a]pyrazin- 1-yl]oxyaceticacid,

[7-Ethyl-3-methyl-8-oxamoyl-6-(2-(2-thienyl)benzyl)pyrrolo[1,2-a]pyrazin-1 -yl]oxyacetic acid,

[7-Ethyl-3-methyl-8-oxamoyl-6-(2-phenylethynylbenzyl)pyrrolo[1,2-a]pyrazin- 1-yl]oxyacetic acid,

[7-Ethyl-3-methyl-8-oxamoyl-6-(2-phenyloxybenzyl) pyrrolo[1,2-a]pyrazin-1-yl]oxyacetic acid,

[7-Ethyl-3-methyl-8-oxamoyl-6-(2-(3-thienyl)benzyl)pyrrolo[1,2-a]pyrazin- 1 -yl]oxyacetic acid,

[7-Ethyl-3-methyl-6-(2-(5-methylthien-2-yl)benzyl)-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyaceticacid,

[7-Ethyl-6-(2-(4-methoxyphenyl)benzyl)-3-methyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl ]oxyacetic acid,

[7-Ethyl-3-methyl-6-(2-(4-methylphenyl)benzyl)-8-oxamoylpyrrolo[1,2-a]pyrazin- 1-yl]oxyacetic acid,

[7-Ethyl-3-methyl-8-oxamoyl-6-(2-(2-phenylethyl)benzyl)pyrrolo[1,2-a]pyrazin-1 -yl]oxyacetic acid,

[6-Benzyl-7-cyclopropyl-3-methyl-8-oxamoylpyrrolo[1,2-a]pyrazin- 1-ylloxyacetic acid,

[7—Cyclopropyl-6-(4-fluorobenzyl)-3-methyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1 -yl]oxyacetic acid,

[6-Benzyl-3-cyclohexyl-7-ethyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyacetic acid,

[6-(2-Biphenylmethyl)-3-cyclohexyl-7-ethyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyacetic acid,

[6-Benzyl-3,7-dimethyl-8-oxamoylpyrrolo [ 1,2-a]pyrazin-1-ylloxyaceticacid,

[7-Ethyl-3-methyl-6-(5-methylthien-2-ylmethyl)-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyacetic acid,

[6-(Benzo[b]thiophen-3-ylmethyl)-7-ethyl-3-methyl-8-oxamoylpyrrolo[1,2-a]pyrazin- 1-yl ]oxyacetic acid,

Sodium [7-ethyl-6-(4-fluorobenzyl)-3-methyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyacetate,

Sodium[7-ethyl-6-(2-(4-fluorophenyl)benzyl)-3-methyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyacetate,

Sodium[7-ethyl-3-methyl-8-oxamoyl-6-(2-(2-thienyl)benzyl)pyrrolo[1,2-a]pyrazin-1-yl]oxyacetate,

Sodium [7-ethyl-3-methyl-8-oxamoyl-6-(2-(3-thienyl)benzyl)pyrrolo[1,2-a]pyrazin-1-yl ]oxyacetate,

and the prodrugs thereof; the parent acids thereof, or theirpharmaceutically acceptable salts; or their solvates.

Most preferred as sPLA₂ inhibitors of the invention are

Methyl[7-ethyl-6-(2-(4-fluorophenyl)benzyl)-3-methyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1 -yl]oxyacetate,

Ethyl[7-ethyl-6-(2-(4-fluorophenyl)benzyl)-3-methyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyacetate,

Morpholinylethyl[7-ethyl-6-(2-(4-fluorophenyl)benzyl)-3-methyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyacetate,

Sodium[7-ethyl-6-(2-(4-fluorophenyl)benzyl)-3-methyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-y)oxyacetate,

Methyl[7-ethyl-3-methyl-8-oxamoyl-6-(2-(2-thienyl)benzyl)pyrrolo[1,2-a]pyrazin-1-yl]oxyacetate, Ethyl[7-ethyl-3-methyl-8-oxamoyl-6-(2-(2-thienyl)benzyl)pyrrolo[1,2-a]pyrazin-1-yl]oxyacetate,

Morpholinylethyl[7-ethyl-3-methyl-8-oxamoyl-6-(2-(2-thienyl)benzyl)pyrrolo[1,2-a]pyrazin-1-yl]oxyacetate,and

Sodium[7-ethyl-3-methyl-8-oxamoyl-6-(2-(2-thienyl)benzyl)pyrrolo[1,2-a]pyrazin-1-yl ]oxyacetate.

The invention also relates to a pharmaceutical composition containing asactive ingredient a compound as described in any one of formula (I) or(II) or (III) or (IV) supra., or as named, supra., or as tabulated inTables 14 to 25, infra., or as described in any one of the Examples,infra.

The invention further relates to a pharmaceutical composition asdescribed in the preceding paragraph, which is for inhibiting sPLA₂.

The invention also relates to a pharmaceutical composition as describedin the preceding paragraph, which is for treatment or prevention ofInflammatory Diseases.

The invention further is also a method of inhibiting sPLA₂ mediatedrelease of fatty acid which comprises contacting sPLA₂ with atherapeutically effective amount of a pyrrolo[1,2-a]pyrazine compound.

The invention is also a method of treating a mammal, including a human,to alleviate the pathological effects of Inflammatory Diseases; whereinthe method comprises administration to said mammal of apyrrolo[1,2-a]pyrazine compound.

The invention further relates to a pyrrolo[1,2-a]pyrazine compound ofdescribed in any one of formula (I) or (II) or (III) or (IV) supra., oras named, supra., or as tabulated in Tables 14 to 25, infra., or asdescribed in any one of the Examples, infra, or a pharmaceuticalformulation containing an effective amount of said compound for use intreatment of Inflammatory Diseases.

The invention also relates to a compound or formulation as described inthe preceding paragraph containing an effective amount of apyrrolo[1,2-a]pyrazine compound for use as an inhibitor for inhibitingsPLA₂ mediated release of fatty acid.

The invention further relates to a pyrrolo[1,2-a]pyrazine sPLA₂inhibitor substantially as hereinbefore described with reference to anyof the Examples.

In the present specification, the term “alkyl” employed alone or incombination with other terms means a straight- or branched chainmonovalent hydrocarbon group having a specified number of carbon atoms.An example of the alkyl includes methyl, ethyl, n-propyl, isopropyl,n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl, n-heptyl,n-octyl, n-nonyl, n-decanyl, n-undecanyl, n-dodecanyl, n-tridecanyl,n-tetradecanyl, n-pentadecanyl, n-hexadecanyl, n-heptadecanyl,n-octadecanyl, n-nonadecanyl, n-eicosanyl and the like.

The term “alkenyl” employed alone or in combination with other terms inthe present specification means a straight- or branched chain monovalenthydrocarbon group having a specified number of carbon atoms and at leastone double bond. An example of the alkenyl includes vinyl, allyl,propenyl crotonyl, isopentenyl, a variety of butenyl isomers and thelike.

The term “alkynyl” used in the present specification means a straight orbranched chain monovalent hydrocarbon group having a specified number ofcarbon atoms and at least one triple bond. The alkynyl may contain (a)double bond(s). An example of the alkynyl includes ethynyl, propynyl,6-heptynyl, 7-octynyl, 8-nonynyl and the like.

The term “carbocyclic group” used in the present specification means agroup derived from a saturated or unsaturated, substituted orunsubstituted 5 to 14 membered, preferably 5 to 10 membered, and morepreferably 5 to 7 membered organic nucleus whose ring forming atoms(other than hydrogen atoms) are solely carbon atoms. A group containingtwo to three of the carbocyclic group is also included in the abovestated group. An example of typical carbocyclic groups includes (f)cycloalkyl (such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cycloheptyl, and cyclooctyl); cycloalkenyl (such as cyclobutylenyl,cyclopentenyl, cyclohexenyl, cycloheptenyl, and cyclooptenyl); phenyl,spiro[5,5]undecanyl, naphthyl, norbornyl, bicycloheptadienyl, tolyl,xylyl, indenyl, stilbenyl, terphenylyl, diphenylethylenyl,phenylcyclohexenyl, acenaphthyl, anthoryl, biphenylyl, bibenzylyl, and aphenylalkylphenyl derivative represented by the formula:

wherein x is an integer from 1 to 8.

The term “spiro[5,5]undecanyl” refers to the group represented by theformula:

Phenyl, cyclohexyl or the like is preferred as a carbocyclic groups inthe R⁴ and R⁵.

The term “heterocyclic group” used in the present specification means agroup derived from monocyclic or polycyclic, saturated or unsaturated,substituted or unsubstituted heterocyclic nucleus having 5 to 14 ringatoms and containing 1 to 3 hetero atoms selected from the groupconsisting of nitrogen atom, oxygen atom, and sulfur atom. An example ofthe heterocyclic group includes pyridyl, pyrroly], pyrrolidinyl,piperidinyl, furyl, benzofuryl, thienyl, benzothienyl, pyrazolyl,imidazolyl, phenylimidazolyl, triazolyl, isoxazolyl oxazolyl, thiazolyl,thiadiazolyl, indolyl, carbazolyl, norharmanyl azaindolyl, benzofuranyl,benzothiophenyl, dibenzofuranyl, dibenzothiophenyl, indazolyl,imidazo[1,2-a]pyridinyl, benzotriazolyl, anthranilyl,1,2-benzisoxazolyl, benzoxazolyl, benzothiazolyl, purinyl, puridinyl,dipyridinyl, phenylpyridinyl, benzylpyridinyl, pyrimidinyl,phenylpyrimidinyl, pyrazinyl, 1,3,5-triazinyl, quinolyl, phthalazinyl,quinazolinyl, quinoxalinyl, morpholino, thiomorpholino, homopiperazinyl,tetrahydrofuranyl, tetrahydropyranyl, oxacanyl, 1,3-dioxolanyl,1,3-dioxanyl, 1,4-dioxanyl, tetrahydrothiopheneyl,pentamethylenesulfadyl, 1,3-dithianyl, 1,4-dithianyl, 1,4-thioxanyl,azetidinyl, hexamethyleneiminium, heptamethyleneiminium, piperazinyl andthe like.

Furyl, thienyl or the like is preferred as a heterocyclic group in theR⁴ and R⁵.

Preferred carbocyclic and heterocyclic groups in R¹ are (g) a grouprepresented by the formula:

wherein R¹³ and R¹⁴ are independently selected from a halogen, C1 to C10alkyl, C1 to C10 alkyloxy, C1 to C10 alkylthio, aryl, heteroaryl, and C1to C10 haloalkyl, a is an oxygen atom or a sulfur atom, L⁵ is —(CH₂)v—,—C═C—, —C≡C—, —O—, or —S—, v is an integer from 0 to 2; α is an oxygenatom or a sulfur atom; β is —CH₂—or —(CH₂)₂—; γ is an oxygen atom or asulfur atom; b is an integer from 0 to 3, d is an integer from 0 to 4;f, p, and w are an integer from 0 to 5; r is an integer from 0 to 7, andu is an integer from 0 to 4. When the above b, d, f, p, r, u, and/or ware 2 or more, a plural number of R¹³ or R¹⁴ may be different from oneanother. When R¹³ is a substituent on the naphthyl group, thesubstituent may be substituted at any arbitrary position on the naphthylgroup. A more preferable example includes (h) a group represented by theformula:

wherein R¹³, R¹⁴, α, β; and γ are the same as defined above, L⁶ is abond, —CH₂—, —C═C—, —C ≡C—, —O—, or —S— and γ is 0 or 1. When R¹³ is asubstituent on the naphthyl group, the substituent may be substituted atany arbitrary position on the naphthyl group.

The “pyrrolo[1,2-a]pyrazine nucleus” is represented by the followingstructural formula together its numerical ring position for substituentplacement:

The term “non-interfering substituent” in the present specificationmeans a group suitable for substitution at position 3 and 4 on thepyrrolo[1,2-a]pyrazine nucleus represented by the formula (I) as well asa group suitable for substitution of the above described “carbocyclicgroup” and “heterocyclic group”. An example of the non-interferingsubstituents includes C1 to C8 alkyl, C2 to C8 alkenyl, C2 to C8alkynyl, C7 to C12 aralkyl (such as benzyl and phenethyl), C7 to C12alkaryl, C2 to C8 alkenyloxy, C2 to C8 alkynyloxy, C3 to C8 cycloalkyl,C3 to C8 cycloalkenyl, phenyl, tolyl, xylyl, biphenylyl, C1 to C8alkyloxy, C2 to C12 alkyloxyalkyl (such as methyloxymethyl,ethyloxymethyl, methyloxyethyl, and ethyloxyethyl), C2 to C12alkyloxyalkyloxy (such as methyloxymethyloxy and methyloxyethyloxy), C2to C12 alkylcarbonyl (such as methylcarbonyl and ethylcarbonyl), C2 toC12 alkylcarbonylamino (such as methylcarbonylamino andethylcarbonylamino), C2 to C12 alkyloxyamino (such as methyloxyamino andethyloxyamino), C2 to C12 alkyloxyaminocarbonyl (such asmethyloxyaminocarbonyl and ethyloxyaminocarbonyl), C1 to C12 alkylamino(such as methylamino, ethylamino, dimethylamino, and ethylmethylamino),C1 to C6 alkylthio, C2 to C12 alkylthiocarbonyl (such asmethylthiocarbonyl and ethylthiocarbonyl), C1 to C8 alkylsulfinyl (suchas methylsulfinyl and ethylsulfinyl), C1 to C8 alkylsulfonyl (such asmethylsulfonyl and ethylsulfonyl), C2 to C8 haloalkyloxy (such as2-chloroethyloxy and 2-bromoethyloxy), C1 to C8 haloalkylsulfonyl (suchas chloromethylsulfonyl and bromomethylsulfonyl), C2 to C8 haloalkyl, C1to C8 hydroxyalkyl (such as hydroxymethyl and hydroxyethyl), —C(O)O(C1to C8 alkyl) (such as methyloxycarbonyl and ethyloxycarbonyl,—(CH₂)z—O—(C1 to C8 alkyl), benzyloxy, aryloxy (such as phenyloxy),arylthio (such as phenylthio), —(CONHSO₂R²⁵), —CHO, amino, amidino,halogen, carbamyl, carboxyl, carbalkyloxy, —(CH₂)z—COOH (such ascarboxymethyl, carboxyethyl, and carboxypropyl), cyano, cyanoguanidino,guanidino, hydrazido, hydrazino, hydroxy, hydroxyamino, nitro,phosphono, —SO₃H, thioacetal thiocarbonyl, carbonyl, carbocyclic groups,heterocyclic groups and the like wherein z is an integer from 1 to 8 andR²⁵ is C1 to C6 alkyl or aryl. These groups may be substituted by atleast one substituent selected from the group consisting of C1 to C6alkyl, C1 to C6 alkyloxy, C2 to C6 haloalkyloxy, C1 to C6 haloalkyl, andhalogens.

Preferable are halogens, C1 to C6 alkyl, C1 to C6 alkyloxy, C1 to C6alkylthio, and C1 to C6 haloalkyl as the “non-interfering substituent”in the R¹. More preferable are halogens, C1 to C3 alkyl, C1 to C3alkyloxy, C1 to C3 alkylthio, and C1 to C3 haloalkyl.

Preferable are (i) C1 to C6 alkyl, aralkyl, C1 to C6 alkyloxy, C1 to C6alkylthio, C1 to C6 hydroxyalkyl, C2 to C6 haloalkyloxy, halogens,carboxy, C1 to C6 alkyloxycarbonyl, aryloxy, arylthio, carbocyclicgroups, and heterocyclic groups as the “non-interfering substituents” inthe R⁴, R⁵, R¹⁰, and R¹¹. More preferable are (0) C1 to C6 alkyl,aralkyl, carboxy, C1 to C6 hydroxyalkyl, phenyl, and C1 to C6alkyloxycarbonyl.

The term “halogen” in the present specification means fluorine,chlorine, bromine, and iodine.

The term “cycloalkyl” in the present specification means a monovalentcyclic hydrocarbon group having a specified number of carbon atoms. Anexample of the cycloalkyl includes cydlopropyl, cyclobutyl, cyclopentyl,cyclohexyl, cycloheptyl, cyclooctyl and the like.

The term “cycloalkenyl” in the present specification means a monovalentcyclic hydrocarbon group having a specified number of carbon atoms andat least one double bond(s). An example of the cycloalkenyl includes1-cyclopropenyl, 2-cyclopropenyl, 1-cyclobutenyl, 2-cyclobutenyl and thelike.

In the present specification, an example of “alkyloxy” includesmethyloxy, ethyloxy, n-propyloxy, isopropyloxy, n-butyloxy, n-pentyloxy,n-hexyloxy and the like.

In the present specification, an example of “alkylthio” includesmethylthio, ethylthio, n-propylthio, isopropylthio, n-butylthio,n-pentylthio, n-hexylthio and the like.

The term “acidic group” in the present specification means an organicgroup functioning as a proton donor capable of hydrogen bonding whenattached to a pyrrolo[1,2-a]pyrazine nucleus through a suitable linkingatom (hereinafter defined as “acid linker”). An example of the acidicgroup includes (k) a group represented by the formula:

wherein R¹⁸ is hydrogen atom, a metal, or C1 to C10 alkyl and each R¹⁹is independently hydrogen atom or C1 to C10 alkyl. Preferable is (1)—COOH, —SO₃H, or P(O)(OH)₂. More preferable is (m)—COOH.

The term “acid linker” in the present specification means a divalentlinking group represented by a symbol —(L²)—, and it functions to join1-position of pyrrolo[1,2-a]pyrazine nucleus to an “acidic group” in thegeneral relationship. An example of it includes (n) a group representedby the formula:

wherein M is —CH₂—, —O—, —N(R²⁴)—, or —S—, and R¹⁶ and R¹⁷ areindependently hydrogen atom, C1 to C10 alkyL aryl, aralkyl, carboxy, orhalogens. Preferable are (o) —O—CH₂—, —S—CH₂, —N(R²⁴)—CH₂—, —CH₂—CH₂—,—O—CH(CH₃)—, or —O—CH((CH₂)₂Ph)— wherein R²⁴ is hydrogen atom or C1 toC6 alkyl and Ph is phenyl. More preferable is (o) —O—CH₂— or —S—CH₂—.

In the present specification, the term “acid linker length” means thenumber of atoms (except for hydrogen atoms) in the shortest chain of alinking group —(L²)—which connects 1-position in pyrrolo[1,2-a]pyrazinenucleus with the “acidic group”. The presence of a carbocyclic ring in—(L²)—counts as the number of atoms approximately equivalent to thecalculated diameter of the carbocyclic ring. Thus, a benzene andcyclohexane ring in the acid linker counts as two atoms in culculatingthe length of —(L²)—. A preferable length is 2 to 3.

A symbol k in the formula (IV) is preferably 1.

The term “haloalkyl” in the present specification means the abovedescribed “alkyl” substituted with the above described “halo(gen” atarbitrary position(s). An example of the halo alkyl includes chloromethyl, trifluoro methyl, 2-chloroethyl, 2-bromoethyl and the like.

The term “hydroxyalkyl” in the present specification means theaforementioned “alkyl” substituted with hydroxy at arbitraryposition(s). An example of the hydroxyalkyl includes hydroxymethyl,2-hydroxyethyl, 3-hydroxypropyl and the like. In this case,hydroxymethyl is preferable.

In the present specification, the term “haloalkyl” in “haloalkyloxy” isthe same as defined above. An example of it includes 2-chloroethyloxy,2,2,2-tritluoroethyloxy, 2-chloroethyloxy and the like.

The term “aryl” in the present specification means a monocyclic orcondensed cyclic aromatic hydrocarbon. An example of the aryl includesphenyl, 1-naphthyl, 2-naphthyl, anthryl and the like. Particularly,phenyl, and 1-naphthyl are preferred.

The term “aralkyl” in the present specification means a group whereinthe aforementioned “alkyl” is substituted with the above-mentioned“aryl”. Such aryl may have a bond at any substitutable position . Anexample of it includes benzyl, phenethyl, phenylpropyl (such as3-phenylpropyl), naphthylmethyl (such as 1-naphthylmethyl) and the like.

The term, “group containing 1 to 4 non-hydrogen atoms” refers torelatively small groups which form substituents at the 7 position of thepyrrolo[1,2-a]pyrazine nucleus, said groups may contain non-hydrogenatoms alone, or non-hydrogen atoms plus hydrogen atoms as required tosatisfy the unsubstituted valence of the non-hydrogen atoms, forexample; (i) groups absent hydrogen which contain no more than 4non-hydrogen atoms such as —CF₃, —Cl, —Br, —NO, —CN, —SO₃; and (ii)groups having hydrogen atoms which contain less than 4 non-hydrogenatoms such as —CH₃, —C₂H₅, —CH═CH₂, —CH(CH₃)₂,and cyclopropyl.

An example of the “alkyloxycarbonyl” in the present specificationincludes methyloxycarbonyl, ethyloxycarbonyl, n-propyloxycarbonyl andthe like.

A group of preferable substituents as the R¹ to R⁵ and the R^(A) of thecompound represented by the formula (I) will be shown in items (A) to(W). Items (f) to (m) are the same group as described above.

As the R¹, (A): —(L¹)—R⁶, (B): —(CH₂)₁₋₂—(g), (C): —(CH₂),₂—(g), and(D): —(CH,)₁₋₂—(h) are preferred.

As the R², (E): hydrogen atom, halogen, C1 to C3 alkyl, C3 to C4cycloalkyl, or C1 to C3 alkyloxy; and (F): C1 to C3 alkyl or C3 to C4cycloalkyl are preferred.

As the R^(A), (G): —C(═O)—C(═O)—NH₂, —CH₂C(═O)—NH₂, or —CH₂C(═O) —NHNH₂;and (H): —C(═O)—C(═O)—NH₂ are preferred.

As the R³, (I): —(n)—(k), (J): —(n)—(I), (K): —(n)—(m), (L): —(o)—(k),(M): —(o)—(l), (N): —(o)—(m), (O): —(p)—(k), (P): —(p)—(l), and (Q):—(p)—(m) are preferred.

As the R⁴, (R): hydrogen atom or non-interfering substituent, (S):hydrogen atom or (i), and (T): hydrogen atom or 0) are preferred.

As the R⁵, (U): hydrogen atom or (i), (V): hydrogen atom or (j), and(W): hydrogen atom are preferred.

A preferred group of compounds represented by the formula (I) will beshown hereinafter.

(R¹,R²,R^(A),R⁴,R⁵)═(A,E,G,R,U), (A,E,G,R,V), (A,E,G,R,W), (AE,G,S,U),(A,E,G,S,V),

(A,E,G,S,W), (A,E,G,T,U), (A,E,G,T,V), (A,E,G,T,W), (A,E,H,R,U),(A,E,H,R,V),

(A,E,H,R,W), (A,E,H,S,U), (A,E,H,S,V), (A,E,H,S,W), (A,E,H,T,U),(AE,H,T,V),

(A,E,H,T,W), (A,F,G,R,U), (AF,G,R,V), (AF,G,R,W), (A,F,G,S,U),(A,F,G,S,V),

(A,F,G,S,W), (A,F,G,T,U), (A,F,G,T,V), (AF,G,T,W), (A,F,H,R,U),(A,F,H,R,V),

(A,F,H,R,W), (A,F,H,S,U), (AF,H,S,V), (AF,H,S,W), (AF,H,T,U),(A,F,H,T,V),

(A,F,H,T,W), (B,E,G,R,U), (B,E,G,R,V), (B,E,G,R,W), (B,E,G,S,U),(B,E,G,S,V),

(B,E,G,S,W), (B,E,G,T,U), (B,E,G,T,V), (B,E,G,T,W), (B,E,H,R,U),(B,E,H,R,V),

(B,E,H,R,W), (B,E,H,S,U), (B,E,H,S,V), (B,E,H,S,W), (B,E,H,T,U),(B,E,H,T,V),

(B,E,H,T,W), (B,F,G,R,U), (B,F,G,R,V), (B,F,G,R,W), (B,F,G,S,U),(B,F,G,S,V),

(B,F,G,S,W), (B,F,G,T,U), (B,F,G,T,V), (B,F,G,T,W), (B,F,H,R,U),(B,F,H.R,V),

(B,F,H,R,W), (B,F,H,S,U), (B,F,H,S,V), (B,F,H,S,W), (B,F,H,T,U),(B,F,H,T,V).

(B ,F,H,T,W), (C,E,G,R,U), (C,E,G,R,V), (C,E,G,R,W), (C,E,G,S,U),(C,E,G,S,V),

(C,E,G,S,W), (C,E,G,T,U), (C,E,G,T,V), (C,E,G,T,W), (C,E,H,R,U),(C,E,H,R,V),

(C,E,H,R,W, (C,E,H,S,U), (C,E,H,S,V), (C,E,H,S,W), (C,E,H,T,U),(C,E,H,T,V),

(C,E,H,T,W), (C,F,G,R,U), (C,F,G,R,V), (C,F,G,R,W), (C,F,G,S,U),(C,F,G,S,V),

(C,F,G,S,W), (C,F,G,T,U), (C,F,G,T,V), (C,F,G,T,W), (C,F,H,R,U),(C,F,H,R,V),

(C,F,H,R,W), (C,F,H,S,U), (C,F,H,S,V), (C,F,H,S,W), (C,F,H,T,U),(C,F,H,T,V),

(C,F,H,T,W), (D,E,G,R,U), (D,E,G,R,V), (D,E,G,R,W), (D,E,G,S,U),(D,E,G,S,V),

(D,E,G,S,W), (D,E,G,T,U), (D,E,G,T,V), (D,E,G,T,W), (D,E,H,R,U),(D,E,H,R,V),

(D,E,H,R,W), (D,E,H,S,U), (D,E,H,S,V), (D,E,H,S,W), (D,E,H,T,U),(D,E,H,T,V),

(D,E,H,T,W), (D,F,G,R,U), (D,F,G,R,V), (D,F,G,R,W), (D,F,G,S,U),(D,F,G,S,V),

(D,F,G,S,W), (D,F,G,T,U), (D,F,G,T,V), (D,F,G,T, W), (D,F,H,R,U),(D,F,H,R,V),

(D,F,H,R,W), (D,F,H,S,U), (D,F,H,S,V), (D,F,H,S,W), (D,F,H,T,U),(D,F,H,T,V), and

(D,F,H,T,W).

Preferred embodiments of this invention are compounds wherein R³ is anyone of (I) to (Q) and (R¹,R²,R^(A),R⁴,R⁵) is any one of the abovecombinations.

The term, “Inflammatory Diseases” refers to diseases such asinflammatory bowel disease, sepsis, septic shock, adult respiratorydistress syndrome, pancreatitis, trauma-induced shock, bronchial asthma,allergic rhinitis, rheumatoid arthritis, chronic rheumatism, arterialsclerosis, cereberal hemorrhage, cerebral infarction, cardiac failure,cardiac infarction, psoriasis, cystic fibrosis, stroke, acutebronchitis, chronic bronchitis, acute bronchiolitis, chronicbronchiolitis, osteoarthritis, gout, spondylarthropathris, ankylosingspondylitis, Reiter's syndrome, psoriatic arthropathy, enterapathricspondylitis, Juvenile arthropathy or juvenile ankylosing spondylitis,Reactive arthropathy, infectious or post-infectious arthritis,gonoccocal arthritis, tuberculous arthritis, viral arthritis, fungalarthritis, syphilitic arthritis, Lyme disease, arthritis associated with“vasculitic syndromes”, polyarteritis nodosa, hypersensitivityvasculitis, Luegenec's granulomatosis, polymyalgin rheumatica, jointcell arteritis, calcium crystal deposition arthropathris, pseudo out,non-articular rheumatism, bursitis, tenosynomitis, epicondylitis (tenniselbow), carpal tunnel syndrome, repetitive use injury (typing),miscellaneous forms of arthritis, neuropathic joint disease (charco andjoint), hemarthrosis (hemarthrosic), Henoch-Schonlein Purpura,hypertrophic osteoarthropathy, multicentric reticulohistiocytosis,arthritis associated with certain diseases, surcoilosis,hemochromatosis, sickle cell disease and other hemoglobinopathries,hyperlipoproteineimia, hypogammaglobulinemia, hyperparathyroidism,acromegaly, familial Mediterranean fever, Behat's Disease, systemiclupus erythrematosis, or relapsing polychondritis and related diseaseswhich comprises administering to a mammal in need of such treatment atherapeutically effective amount of the compound of formula I in anamount sufficient to inhibit sPLA₂ mediated release of fatty acid and tothereby inhibit or prevent the arachidonic acid cascade and itsdeleterious products.

The terms, “mammal” and “mammalian” include human.

The term “solvate” includes, for example, solvates with organicsolvents, hydrates, and the like.

The compounds of the invention represented by the general formula (I)can be synthesized in accordance with the following methods A to I.

wherein R², R⁴, R⁵, R⁶ , R²², X, Y, and L⁴ are as defined above; R²⁶R²⁷, and R²⁸ are C1 to C3 alkyl; Hal is a halogen, and Met is an alkalimetal.

(Step 1)

The present step is the one for constructing pyrrolo[1,2-a]pyrazinering, and it may be conducted in accordance with a process described inJ. Chem. Soc., Perkin Trans. 1, 1990, 311-314 (The disclosure of whichare incorporated herein by reference).

(Step 2)

The present step is the one for transforming the ketone at 1-positioninto an alkyloxy group. To the compound (VII) is added a halogenatingagent such as phosphorus oxychloride, phenylphosphonic dichloride andthe like, and the resulting mixture is refluxed for 1 to 8 hours,preferably 3 to 5 hours. The resulting compound is dissolved in analcohol (for example, methanol, ethanol, and n-propanol), an alkalimetal compound of C1 to C3 alcohol (for example, sodium methoxide, andsodium ethoxide), sodium p-toluenesulfinate and the like are added tothe solution, and the mixture is stirred at 70° C. to 120° C.,preferably 80° C. to 100° C. for 5 to 36 hours, preferably 12 to 24hours. When the resulting product is subjected to a usual work-up, thecompound (VIII) can be obtained.

(Step 3)

The present step is the one for introducing a substituent to 6-positionof pyrrolo[1,2-a]pyrazine, and it may be carried out by Friedel—Craftsreaction. The compound (VIII) is dissolved in a solvent such as1,2-dichloroethane, methylene chloride and the like, R⁶COHal and Lewisacid (for example, AlCl₃, SbF₅, BF₃ and the like) are added gradually tothe solution at −78° C. to 10° C., preferably −20° C. to ice-cooling,and the resulting mixture is stirred at −10° C. to 10° C., preferably 0°C. to 10° C. for 5 to 30 minutes, preferably 10 to 20 minutes.Alternatively, the reaction may be carried out in such that the compound(VIII) is dissolved in R⁶COHal without using any solvents, and then, thestep is continued in accordance with the same manner as that describedabove. When the resulting product is subjected to a usual work-up, thecompound (IX) can be obtained (see J. Med. Chem., 39, 3636-58 (1996).The disclosure of which are incorporated herein by reference.)

(Step 4)

The present step is the one for reducing the carbonyl group at6-position of pyrrolo[1,2-a]pyrazine to transform the same intomethylene. Lewis acid (for example, AlCl₃ and the like) is dissolved ina solvent such as methylene chloride, tetrahydrofuran and the like, areducing agent such as boron-t-butylamine complex, sodium borohydrideand the like is added to the solution at −20° C. to 10° C., preferablyunder ice-cooling, and the resulting mixture is stirred for 5 to 30minutes, preferably 10 to 20 minutes. The compound (IX) dissolved inmethylene chloride, tetrahydrofuran and the like is added to thereaction mixture at −20° C. to 10° C., preferably under ice-cooling, theresulting mixture is stirred preferably for 20 to 30 minutes, andfurther the stir is continued at 15° C. to 40° C., preferably 20° C. to30° C. for 1 to 5 hours, preferably 2 to 3 hours. When the resultingproduct is subjected to a usual work-up, the compound (X) can beobtained (see J. Med. Chem., 39, 3635-58 (1996). It is to be noted thatThe disclosure of which are incorporated herein by reference.)

(Step 5)

The present step is the one for transforming the alkyloxy group intoketone. An acid such as concentrated hydrochloric acid and the like isadded to the compound (X), and the mixture is stirred at 80° C. to 150°C., preferably 100° C. to 120° C. for 1 to 5 hours preferably 2 to 3hours. When the resulting product is subjected to a usual work-up, thecompound (XI) can be obtained.

(Step 6)

The present step is the one for transforming the ketone at 1-positioninto a halogen. A halogenating agent such as phosphorus oxychloride,phenylphosphonic dichloride and the like is added to the compound (XI),and the mixture is refluxed for 1 to 8 hours, preferably 3 to 5 hours.When the resulting product is subjected to an ordinary work-up, thecompound (XII) can be obtained.

(Step 7)

The present step is the one for transforming the halogen at 1-positioninto (—L⁴—R²²). To a suspension of R²²—L⁴—H and an alkali metal compoundsuch as sodium and the like are added the compound (XII) and sodiump-toluenesulfinate or the like, and the mixture is stirred at 70° C. to120° C., preferably 80° C. to 100° C. for 5 to 36 hours, preferably 12to 24 hours. When the resulting product is subjected to an ordinarywork-up, the compound (XIII) can be obtained.

(Step 8)

The present step is the one for introducing a substituent to 8-position.The compound (XIII) is dissolved in a solvent such as1,2-dichloroethane, tetrahydrofuran and the like, Hal—C(═X)—C(═X)—Hal(for example, oxalyl chloride) and a base such as N-methylmorpholine,triethylamine and the like are added to the solution, and the mixture isstirred at 30° C. to 70° C., preferably 40° C. to 60° C. for 1 to 10hours, preferably 3 to 6 hours. The reaction mixture is poured into coldaqueous ammonia, and the resulting mixture is stirred for 5 to 30minutes, preferably 10 to 20 minutes. When the resulting product issubjected to an ordinary work-up, the compound (XV) can be obtained.

wherein R², R⁴, R⁶, R²², R28, L⁴, X, Y, and Hal are as defined above,and R⁵ is hydrogen.

(Step 1)

The present step is the one for constructing pyrazine ring, and it maybe carried out in accordance with the process described in J. Am. Chem.Soc., 74, 1580-84 (1952). (The disclosure of which are incorporatedherein by reference.)

(Step 2)

The present step may be carried out in accordance with the same manneras that of the method A—step 2.

(Step 3)

The present step is the one for constructing pyrrolo[1,2-a]pyrazinering. A mixture of the compound (XIX) and Hal—CH₂—C(═O)—R² is stirred at40° C. to 90° C., preferably 50° C to 70° C. for 3 to 36 hours,preferably 12 to 24 hours to obtain a quaternary salt. The resultingquaternary salt is dissolved in a solvent such as 1,2-dichloroethane,acetonitrile and the like, a base such as1,8-diazabicyclo[5,4,0]-undec-7-ene(DBU), triethylamine and the like isadded to the solution, and the mixture is stirred at 40° C. to 90° C.,preferably 50° C. to 70° C. for 3 to 36 hours, preferably 12 to 24hours. When the resulting product is subjected to a usual work-up, thecompound (XXI) can be obtained.

(Step 4)

The present step may be carried out in accordance with the same manneras that of the method A—step 3.

(Step 5)

The present step may be carried out in accordance with the same manneras that of the method A—steps 4 to 8.

wherein R⁴, R⁵, and Hal are as defined above, and Boc ist-butoxycarbonyl.

(Step 1)

The present step is the one for conducting condensation reaction of thecompound (XXIII) and the compound (XXIV). The compound (XXIII) isdissolved in a solvent such as tetrahydrofuran, dichloromethane,acetonitrile and the like, the compound (XXIV) and a condensation agentsuch as N,N-dicyclohexylcarbodiimide (DCC),1-ethyl-3—(3-dimethylaminopropyl)carbodiimide (WSCD),N,N-dicarbonylimidazole, 2-halo-1-methylpyridinium iodide, di-2-pyridylcarbonate, 1,1′-oxalyldiimidazole and the like are added to thesolution, and the resulting mixture is reacted at −20° C. to 80° C.,preferably 0° C. to 40° C. for 1 to 30 hours, preferably 3 to 20 hoursto obtain the compound (XXV).

(Step 2)

The present step is the one for effecting oxidation of hydroxyl groupand ring closure reaction.

The oxidation reaction may be carried out in accordance with a mannerapplied generally. In this respect, the following four types ofoxidation reaction are particularly preferred.

i) PCC Oxidation (The compound (XXV) is dissolved in a solvent such asdichloromethane and the like, pyridinium chlorochromate (PCC) is addedto the solution, and the mixture is allowed to react at −20° C. to 60°C., preferably 0° C. to 40° C. for 1 to 30 hours, preferably 3 to 20hours, to give an oxidized product.) (see Tetrahedron Lett., 2647-2650(1975))

ii) Swern Oxidation (Dichioromethane is cooled to −78° C., oxalylchloride, dimethyl sulfoxide, and the compound (XXV) are addedsuccessively to the solvent. The mixture is allowed to warm to −45° C.to 0° C., the mixture is allowed to react for 1 to 30 hours, preferably1 to 10 hours. When the resulting product is subjected to a usualwork-up, a aimed compound can be prepared.) (see J. Org. Chem., 43,2480-2482 (1978))

iii) Dess-Martin Oxidation (A solution of Dess-Martin reagent indimethyl sulfoxide or the like is allowed to react with compound (XXV)in a solvent such as tetrahydrofuran.) (see J. Org. Chem., 48, 4155-4156(1983))

iv) Oxidation by Halogen Oxoacid (The compound (XXV) is allowed to reactwith an oxidizing agent such as halogen oxoacid and the like in thepresence of 2,2,6,6-tetramethyl-1-piperizinyloxy (TEMPO) according tothe process described in a literary document (J. Org. Chem., 52,2559-2562 (1987)), whereby the compound can be prepared. In stead ofTEMPO, 4-acetylamino-2,2,6,6-tetramethyl-l-piperidinyloxy,4-benzoyloxy-2,2,6,6-tetramethyl-1-piperidinyloxy,4-cyano-2,2,6,6-tetramethyl-1-piperidinyloxy or the like may be used. Asthe halogen oxoacid, sodium hypochlorite, sodium hypobromite, sodiumbromite or the like is used. As the solvent, ethyl acetate,acetonitrile, dichloromethane or the like may be used.)

In ring closure reaction, the oxidized product prepared in accordancewith the above step is dissolved in a solvent such as toluene, ethylacetate, chloroform and the like, and the solution is allowed to reactat −10° C. to 80° C., preferably 0° C. to 40° C. for 1 to 30 hours,preferably 5 to 20 hours, whereby the compound (XXVI) can be obtained.In the case where progress of the reaction is slow, it is sufficient toadd a catalytic amount of a suitable acid (for example,p-toluenesulfonic acid and the like) to the solution.

(Step 3)

The present step is the one for deprotecting Boc group. The compound(XXVI) is dissolved in a solvent such as dichloromethane, ethyl acetate,toluene and the like, a mineral acid (for example, HCl, HBr, HI and thelike) or an organic acid (for example, trifluoroacetic acid,camphorsulfonic acid and the like) is added to the solution, and themixture is allowed to react at 0° C. to 100° C., preferably 20° C. to100° C. for 1 to 20 hours, preferably 3 to 10 hours, whereby thecompound (XXVII) can be prepared.

(Step 4)

The present step is the one for conducting dehydrogenation reaction. Thecompound (XXVII) is dissolved in a solvent such as decaline, quinoline,naphthalene and the like, Pd, Pt, Rh, Ni, S, or Se is added to thesolution, and the mixture is allowed to react at 100° C. to 350° C. for2 to 5 hours, whereby the compound (XVIII) can be obtained. In the casewhen a hydrogen receptor such as cyclohexene, maleic acid, and the likeis allowed to exist in the reaction system, it is sufficient to be areaction temperature of 100° C. to 150° C.

wherein R⁴ and R⁵ are as defined above.

(Step 1)

The present step may be carried out in accordance with the same manneras that of the method C—step 1.

(Step 2)

The present step may be carried out in accordance with the same manneras that of the oxidation step in the method C—step 2.

(Step 3)

The present step is the one for oxidizing methylene to form ketone. Thecompound (XXX) is dissolved in dichloromethane-methanol, ethyl acetateor the like, and ozone gas is bubbled through the solution at −78° C. to0° C., preferably −78° C. to −30° C. After 5 minutes to 1 hour, dimethylsulfide or triphenylphosphine is added to the resulting mixture, and themixture is allowed to react at 0° C. to 60° C., preferably 10° C. to 40°C. for 1 to 2 hours, whereby the compound (XXXI) can be obtained.

(Step 4)

The present step is the one for effecting ring closure reaction. Thecompound (XXXI) is dissolved in a solvent such as ethanol and the like,ammonium acetate is added to the solution, and the mixture is refluxedfor 5 minutes to 1 hour, whereby the compound (XVIII) can be prepared.

wherein R², R⁴, R⁵, R²², L⁴, X and Y are as defined above, R²⁹ is arylor heteroaryl having a leaving group such as halogen, triilate, R³⁰ isaryl or heteroaryl substituted with aryl, heteroaryl, substituted vinyl,substituted acetylene, alkyl, aryloxy and the like.

(Step 1)

The present step is a step of a carbon-carbon bond forming reaction bySuzuki reaction or Sonogashira reaction using a palladium catalyst. Bythe present reaction, the compound (XXXII) is converted into thecompound (XXXIII) in accordance with the methods described in Syn.Commun., 11, 513 (1981) (The disclosure of which are incorporated hereinby reference), Tetrahedron Lett., 4467 (1975) (The disclosure of whichare incorporated herein by reference) and the like.

Compound (XXXII) is reacted with optionally substituted aryl oroptionally substituted heteroaryl having a B(OH)₂ (otherwise B(Et)₂)group such as phenylboronic acid in a solvent such as dimethylformamide,toluene, xylene, benzene, tetrahydrofuran etc. in the presence of apalladium catalyst (e.g., Pd(Ph₃,P)₄) and a base (e.g., potassiumcarbonate, calcium carbonate, triethylamine, sodium methoxide etc.) togive the desired compound (XXXIII). This reaction is carried out at 0 to100° C., preferably 0 to 80° C. This reaction is completed for 5 to 50hours, preferably 15 to 30 hours. When optionally substituted aryl oroptionally substituted heteroaryl has a substituent(s) interfering thisreaction, the substituent(s) can previously be protected in accordancewith a method of “Protective Groups in Organic Synthesis ” ( Theodora W.Green (John Wiley & Sons)) and then deprotected at an appropriate step.

Compound (XXXII) is reacted with optionally substituted aryl oroptionally substituted heteroaryl having an ethynyl group such asethynylbenzene in a solvent such as dimethylformamide, toluene, xylene,benzene, tetrahydrofuran etc. in the presence of a palladium catalyst(e.g., Pd(Ph₃P)₂Cl₂), a divalent copper reagent (e.g., CuI), and anorganic base (e.g., triethylamine, and diisopropylethylamine) to give adesired compound (XXXIII). This reaction is carried out at 0 to 100° C.,preferably 20 to 80° C. This reaction is completed for 3 to 30 hours,preferably 10 to 20 hours. When optionally substituted aryl oroptionally substituted heteroaryl has a substituent(s) interfering thisreaction, the substituent(s) can previously be protected in accordancewith a method of “Protective Groups in Organic Synthesis” ( Theodora W.Green (John Wiley & Sons)), and then deprotected at an appropriate step.

In case that R³⁰ is aryl or heteroaryl substituted with aryloxy, thecompound (XXXII) is dissolved in a solvent such as pyridine, and thencooper (II) oxide, a base (for example, potassium carbonate) andsubstituted phenols are added, and the resulting mixture was stirred at10 to 150° C., preferably 100 to 150° C., for 1 to 24 hours, preferably5 to 10 hours. The compound (XXXIII) is obtained by the usual work-up.

(Step 2)

The present step can be carried out in the same manner as thosedescribed in step 6 to 8 of Method A

wherein R², R⁴, R⁵, R⁶, R²², X and Y are as defined above.

When L⁴ is CH₂S in Method A, (XXXVII) can also be synthesized by MethodF.

(Step 1)

The present step is a step wherein the ketone group at C1-position isconverted into thioketone. The reaction may be conducted in accordancewith the method described in Monatsh chem, 126, 747 (1995) (Thedisclosure of which are incorporated herein by reference). The compound(XI) is dissolved in a solvent such as pyridine, and the resultingmixture is stirred with phosphorus pentasulfide at 10° C. to 150° C.,preferably 100 to 150° C., for 1 to 5 hours, preferably 2 to 3 hours.The compound (XXXV) is obtained by the usual work-up. This step can alsobe conducted by reacting with compound (XI) and Lawesson reagent in asolvent such as tetrahydrofuran, dimethylformamide at 10 to 150° C.,preferably 50 to 100° C., for 1 to 5 hours, preferably 2 to 3 hours.

(Step 2)

The present step is a step wherein the thioketone group at C1-positionis converted into iminosulfide group.

The compound (XXXV) is dissolved in a solvent such as tetrahydrofuran,dimethylformamide, R²²CH₂X (for example, bromoacetic acid methyl ester)and a base (for example, potassium carbonate) are added, and theresulting mixture is stirred at 0 to 100° C., preferably 10 to 50° C.,for 1 to 5 hours, preferably 1 to 2 hours. The compound (XXXVI) isobtained by the usual work-up.

(Step 3)

The present step can be carried out in the same manner as that describedin step 8 of Method A.

wherein R², R⁴, R⁶, R²², R²⁸, L⁴, Hal, X and Y are as defined above,R³¹L is C1-C3 alkyl or aryl.

(Step 1)

The present step is a step wherein a substituent is introduced toC4-position of pyrrolo[1,2-a]pyrazine without any substituent atC4-position. The compound (XXXVIII) is dissolved in a solvent such asdiethyl ether, tetrahydrofuran, an alkyllithium (for example,methyllithium, n-butyllithium) was added at −78 to 10° C., preferably−30° C. to ice-cooling, and then the resulting mixture is stirred for 15minutes to 1 hour, preferably 15 to 30 minutes. R⁶—CHO is added to theabove mixture and the mixture was stirred further 15 minutes to 1 hour,preferably 15 to 30 minutes. The compound (XXXIX) is obtained by theusual work-up.

(Step 2)

The present step is a step wherein the hydroxyl group at C4-position ofpyrrolo[1,2-a]pyrazine is reduced, and converted into methylene group.The reaction can be conducted in accordance with the method described inTetrahedron, 51, 11043 (1995) (The disclosure of which are incorporatedherein by reference). Alternatively, the reaction may be conducted inaccordance with the above step 4 of Method A, a catalytic hydrogenationmethod by using a reduction catalyst such as palladium-carbon and sourceof hydrogen such as hydrogen gas, ammonium formate (refer to Synth.Commun., 22, 2673 (1992), The disclosure of which are incorporatedherein by reference), a method by using samarium iodide (refer toTetrahedron Lett., 30, 2945 (1989), The disclosure of which areincorporated herein by reference) and the like.

(Step 3)

The present step can be carried out in the same manner as that describedin step 5 of Method A

(Step 4)

The present step can be carried out in the same manner as that describedin step 6 of Method A.

(Step 5)

The present step is a step wherein chloro group at C1-position ofpyrrolo[1,2-a]pyrazine is converted to sulfonyl group. The compound(XLII) is dissolved in an alcoholic solvent such as ethanol or dimethylsulfoxide, a sulfinate salt (for example, sodium p-toluenesulfinate) wasadded, and then the resulting mixture is stirred at 10 to 150° C.,preferably 50 to 100° C., for 1 to 18 hours, preferably 3 to 8 hours.The catalytic amount of acid (for example, hydrochloric acid) may beadded preferably. The compound (XLIII) is obtained by the usual work-up.

(Step 6)

The present step can be carried out in the same manner as that describedin step 7 of Method A.

(Step 7)

The present step can be carried out in the same manner as that describedin step 8 of Method A

wherein R¹, R², R²² , L⁴, X, Y and Hal are as defined above, R³² and R³⁵are C1-C3 alkyl, R³³ is lower alkyl, or a group which forms1,3-dioxolane ring or 1,3-dioxane ring together with the adjacent oxygenatoms, R³⁴ is hydrogen atom, C1-C6 alkyl, C7-C12 aralkyl, C1-C6alkyloxy, C1-C6 alkylthio, C1-C6 hydroxyalkyl, C2-C6 haloalkyloxy,halogen, carboxy, C1-C6 alkyloxycarbonyl, aryloxy, arylthio, acarbocyclic group or a heterocyclic group, Met is metal.

(Step 1)

The compound (XLVI) is dissolved in a solvent such as dimethylfomamide,an aikyl halide derivative (for example, bromoacetaldehydeethyleneacetal and the like) and a base (for example, potassiumcarbonate, potassium t-butoxide, sodium hydride and the like) are added, and then the resulting mixture is stirred at 10 to 80° C., preferably20 to 60° C., for 3 to 80 hours, preferably 5 to 70 hours. The compound(XLVII) is obtained by the usual work-up.

(Step 2)

The present step is a step of decarboxylation reaction. The compound(XLVII) is dissolved in a solvent such as dimethyl sulfoxide, a reagentsuch as potassium acetate, sodium acetate are added, and then theresulting mixture is stirred at 20 to 200° C., preferably 100 to 180°C., for 1 to 20 hours, preferably 3 to 15 hours. The compound (XLVIII)is obtained by the usual work-up.

(Step 3)

The present step is a step of addition reaction of alkyl metal reagentto nitrile group. A solution of the compound (XLVIII) in diethyl ether,tetrahydrofuran, dimethoxyethane or the like is added to Grignardreagent (R¹MgHal, Hal is halogen) or a solution of R¹Li in diethylether, tetrahydrofuran or dimethoxyethane at −20 to 30° C., and themixture is stirred at 0 to 70° C, preferably 20 to 60° C, for 1 to 20hours, preferably 2 to 10 hours. The compound (IL) is obtained by theusual work-up by using an acid such as diluted sulfuric acid.

(Step 4)

The present step is a step for constructing pyrrole ring. The compound(IL) is dissolved in a solvent such as tetrahydrofuran, substitutedallylamine and a catalytic amount of an acid (for example, 1Nhydrochloric acid) are added, and then the mixture is stirred at 0 to100° C, preferably 0 to 50° C, for 1 to 5 hours, preferably 1 to 2 . Thecompound (L) is obtained by the usual work-up. Alternatively, thecompound (IL) is converted into ketoaldehyde derivative by hydrolysis ofacetal portion using an acid such as hydrochloric acid in a solvent suchas tetrahydrofuran. Subsequently, the mixture is treated withsubstituted abylamine in a suitable solvent at 0 to 100° C., preferably0 to 50° C., for 1 to 5 hours, preferably 1 to 2 hours to obtain thecompound (L).

(Step 5)

The present step is a step for introducing alkoxycarbonyl group topyrrole ring. The reaction can be carried out as described in step 3 ofMethod A by using chlorocarbonate. Alternatively, the compound (L) isconverted into trichloroacetyl form by stirring it with trichloroacetylchloride in a solvent such as tetrahydrofuran at 0 to 100° C.,preferably 10 to 40° C., for 1 to 5 hours, preferably 1 to 2 hours.Subsequently, in a suitable alcohol, the mixture is treated with metalalkoxide of the same alcohol at 0 to 100° C., preferably 10 to 60° C.,for 1 to 5 hours, preferably 1 to 2 hours to obtain the compound (LI).

(Step 6)

The present step is a step for constructing pyrrolomorpholine ring byiodo lactonization reaction. The compound (LI) is dissolved in a solventsuch as acetonitrile, iodine was added, and the mixture is stirred at 0to 50° C., preferably 10 to 30° C., for 1 to 10 hours, preferably 1 to 3hours. The compound (LII) is obtained by the usual work-up.

(Step 7)

The present step is a step for forming double bond by eliminating HI.The compound (LII) is dissolved in a solvent such as toluene,acetonitrile, tetrahydrofuran, a base such as1,8-diazabicyclo[5.4.0]-7-undecene is added, and the mixture is stirredat 0 to 100° C., preferably 20 to 80° C., for 1 to 5 hours, preferably 1to 3 hours. The compound (LIII) is obtained by the usual work-up.

(Step 8)

The present step is a step for constructing pyrrolo[1,2-a]pyrazine ring,and can be conducted in accordance with the method described in J. Org.Chem., 53, 4650 (1988) (The disclosure of which are incorporated hereinby reference). The compound (LIII) is dissolved in an alcoholic solventor a solvent such as acetonitrile, tetrahydrofuran, a source of ammoniasuch as ammonium acetate is added, and the mixture is stirred at 0 to100° C., preferably 20 to 80° C., for 3 to 24 hours, preferably 5 to 18hours. The compound (LIV) is obtained by the usual work-up.

(Step 9)

The present step can be carried out in the same manner as thosedescribed in steps 6 to 8 of Method A.

wherein R², R⁶, R²², R²⁶, R³⁴, L⁴, X, Y and Hal are as defined above.

(Step 1)

The present step can be carried out in the same manner as that describedin step 3 of Method A

(Step 2)

The present step can be carried out in the same manner as that describedin step 4 of Method A

(Step 3)

The present step is a step of allylation of nitrogen at N1-position ofpyrrole. The compound (LVII) is dissolved in a solvent such astetrahydrofuran, dimethylformamide, allyl halide derivative and a base(for example, sodium hydride, potassium carbonate) is added, and themixture is stirred at 0 to 100° C., preferably 0 to 50° C., for 1 to 10hours, preferably 1 to 3 hours. The compound (LVIII) is obtained by theusual work-up.

(Step 4)

The present step can be carried out in the same manner as thosedescribed in steps 6 to 9 of Method H.

Where a compound of the present invention has an acidic or basicfunctional group, a variety of salts each having higher water solubilityand more physiologically suitable properties than those of the originalcompound can be formed. An example of typical pharmaceuticallyacceptable salts includes salts with alkali metal and alkaline earthmetal such as lithium, sodium, potassium, magnesium, aluminum and thelike, but it is to be noted that such pharmaceutically acceptable saltsare not limited thereto. A salt is easily manufactured from a free acidby either treating an acid in a solution with a base, or allowing anacid to be in contact with an ion exchange resin. Addition salts of thecompounds according to the present invention with relatively non-toxicinorganic bases and organic bases, for example, amine cation, ammonium,and quaternary ammonium derived from nitrogenous bases having a basicitysufficient for forming a salt of the compounds of the present inventionare included in the definition of “pharmaceutically acceptable salts”.(e.g., S. M. Berge et al., “Pharmaceutical Salts,” J. Phar. Sci., 66,1-19 (1977)) Furthermore, basic groups of a compound according to thepresent invention are reacted with a suitable organic or inorganic acidto form salts such as acetates, benzenesulfonates, benzoates,bicarbonates, bisulfates, bitartarate, borates, bromides, camcyrates(phonetic), carbonates, chlorides, clubranates (phonetic), citrates,edetates (phonetic), edicirates (phonetic), estrates (phonetic),ethylates, fluorides, fumarates, gluseptates (phonetic), gluconates,glutamates, glycolialsanyrates (phonetic), hexylresorcinates,hydroxynaphthoates, iodides, isothionates, lactates, lactobionates,laurates, malates, malseates (phonetic), manderates (phonetic),methylates, methylbromides, methylnitrates, methylsulfates, mucates,napcylates (phonetic), nitrates, oleates, oxarates, palnitates,pantothenates, phosphates, polygalacturonates, salicirates, stearates,subacetates (phonetic), sucinates (phonetic), tanates (phonetic),tartrates, tosylates, trifluoroacetates, trifluoromethanesulfonates,valerates and the like. In case of forming a hydrate, a questionedcompound may be coordinated with a suitable number of water molecules.

In the case where a compound of the present invention has one or more ofchiral center(s), it may exist as an optically active member. Likewise,in the case where a compound contains alkenyl or alkenylene, there is apossibility of cis- and trans-isomers. Mixtures of R- and S-isomers aswell as of cis- and trans-isomers, and mixtures of R- and S-isomerscontaining racemic mixture are included in the scope of the presentinvention. Asymmetric carbon atom may exist also in a substituent suchas alkyl group. All such isomers are included in the present inventiontogether with these mixtures. In the case where a specified streoisomeris desired, either it is manufactured by applying a manner which hasbeen well known by those skilled in the art wherein a starting materialhaving an asymmetrical center which has been previously separated issubjected to stereospecific reaction to the starting material, or it ismanufactured by preparing a mixture of stereoisomers, and thereafterseparating the mixture in accordance with a well-known manner. Forexample, a racemic mixture may be reacted with a single enantiomer ofsome other compound. This changes the racemic form into a mixture ofdiastereomers and diastereomers, because they have different meltingpoints, different boiling points, and different solubilities can beseparated by conventional means, such as crystallization.

Prodrug is a derivative of the compound having a group which can bedecomposed chemically or metabolically, and such prodrug is a compoundaccording to the present invention which becomes pharmaceutically activeby means of solvolysis or by placing the compound in vivo under aphysiological condition. Although a derivative of the compoundsaccording to the present invention exhibits activity in both forms ofacid derivative and basic derivative, acid derivative is moreadvantageous in solubility, tissue affinity, and release control inmammal organism (Bungard, H., Design of Prodrugs, pp. 7-9, 21-24,Elsevier, Amsterdam, 1985). Ester prodrugs are well known (see,Silverman, Richard B, The Organic Chemistry of Drug Design and DrugAction, Chapter 8, New York, N.Y. Academic Press, ISBN 0-12-643730-0)and are a preferred prodrug form for the compounds of this invention andalso for prodrugs used in the method of treating Inflammatory Disease astaught herein. For instance, prodrugs each containing an acid derivativesuch as an ester which is prepared by reacting a basal acid compoundwith a suitable alcohol, or an amide which is prepared by reacting abasal acid compound with a suitable amine are well known by thoseskilled in the art. Simple aliphatic or aromatic esters derived fromacid groups contained in the compounds according to the presentinvention are preferable prodrugs. Particularly preferred esters asprodrugs are methyl, ethyl, propyl, isopropyl n-butyl, isobutyl,tert-butyl morpholinoethyl, and N,N-diethylglycolamido.

Methyl ester prodrugs may be prepared by reaction of the sodium salt ofa compound of Formula (I) (in a medium such as dimethylformamide) withiodo methane (available from Aldrich Chemical Co., Milwaukee, Wis. USA;Item No. 28,956-6).

Ethyl ester prodrugs may be prepared by reaction of the sodium salt of acompound of Formula (I) (in a medium such as dimethylformamide) withiodo ethane (available from Aldrich Chemical Co., Milwaukee, Wis. USA;Item No. 1-778-0).

N,N-diethylglycolamido ester prodrugs may be prepared by reaction of thesodium salt of a compound of Formula (I) (in a medium such asdimethylformamide) with 2-chloro-N,N-diethylacetamide (available fromAldrich Chemical Co., Milwaukee, Wis. USA; Item No. 25,099-6).

Morpholinylethyl ester prodrugs may be prepared by reaction of thesodium salt of a compound of Formula (I) (in a medium such asdimethylformamide) with 4—(2-chloroethyl)morpholine hydrochloride(available from Aldrich Chemical Co., Milwaukee, Wis. USA, Item No.C4,220-3).

Double ester such as (acyloxy)alkyl ester or((alkyloxycarbonyl)oxy)alkyl ester type prodrugs may be optionallymanufactured.

The term “inhibit” means that release of fatty acid started by sPLA₂decreases significantly by the compounds of the present invention fromviewpoint of prevention and treatment of disease. The term“pharmaceutically acceptable” means that carriers, diluents, oradditives are compatible with other ingredients in a formulation and arenot harmful for recipients.

The compounds of the present invention exhibit sPLA₂ inhibiting activityas per the description of the experimental examples which will bedescribed hereinafter. Accordingly, when a curatively effective amountof the compounds represented by the formulae (I), (II), (III), and (IV),the prodrug derivatives thereof, or their pharmaceutically acceptablesalts, or their solvates is administered to any of mammals (includinghuman being), it functions effectively as a curative medicine fordiseases of septic shock, adult respiratory distress syndrome,pancreatitis, injury, bronchial asthma, allergic rhinitis, chronicrheumatism, arterial sclerosis, cerebral hemorrhage, cerebralinfarction, inflammatory colitis, mange, cardiac failure, cardiacinfarction.

The compounds of the present invention may be administered to a patientthrough a variety of routes including oral, aerosol, rectal,percutaneous, subcutaneous, intravenous, intramuscular, and nasalroutes. A formulation according to the present invention may bemanufactured by combining (for example, admixing) a curatively effectiveamount of a compound of the present invention with a pharmaceuticallyacceptable carrier or diluent. The formulation of the present inventionmay be manufactured with the use of well-known and easily availableingredients in accordance with a known method.

In case of manufacturing a composition according to the presentinvention, either active ingredients are admixed with a carrier, or theyare diluted with a carrier, or they are contained in a carrier in theform of capsule, sacheier (phonetic), paper, or another container. Incase of functioning a carrier as a diluent, the carrier is a solid,semi-solid, or liquid material which functions as a medium. Accordingly,a formulation according to the present invention may be produced in theform of tablet, pill, powder medicine, intraoral medicine, elixir agent,suspending agent, emulsifier, dissolving agent, syrup agent, aerosolagent (solid in liquid medium), and ointment. Such a formulation maycontain up to 10% of an active compound. It is preferred to prepare acompound according to the present invention prior to administration.

Any suitable carrier which has been well known by those skilled in theart may be used for the formulation. In such formulation, a carrier isin the form of solid, liquid, or a mixture of solid and liquid. Forinstance, a compound of the present invention is dissolved into 4%dextrose/0.5% sodium citrate aqueous solution so as to be 2 mg/mlconcentration for intravenous injection. Solid formulation includespowder, tablet, and capsule. Solid carrier consists of one or more ofmaterial(s) for serving also as fragrant, lubricant, dissolving agent,suspension, binder, tablet disintegrator, capsule. A tablet for oraladministration contains a suitable excipient such as calcium carbonate,sodium carbonate, lactose, calcium phosphate and the like together witha disintegrator such as corn starch, alginic acid and the like and/or abinder such as gelatin, acacia and the like, and a lubricant such asmagnesium stearate, stearic acid, talc and the like.

In a powder medicine, a carrier is a finely pulverized solid which isblended with finely pulverized active ingredients. In a tablet, activeingredients are admixed with a carrier having required binding power ina suitable ratio, and it is solidified in a desired shape and size.Powder medicine and tablet contain about 1 to about 99% by weight of theactive ingredients being novel compounds according to the presentinvention. An example of suitable solid carriers includes magnesiumcarbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin,starch, gelatin, tragacanth gum, methyl cellulose, sodiumcarboxymethylcellulose, low-melting wax, and cocoa butter.

An axenic liquid formulation contains suspending agent, emulsifier,syrup agent, and elixir agent. Active ingredients may be dissolved orsuspended into a pharmaceutically acceptable carrier such as sterilewater, a sterile organic solvent, a mixture thereof and the like. Activeingredients may be dissolved frequently into a suitable organic solventsuch as propylene glycol aqueous solution. When finely pulverized activeingredients are dispersed into aqueous starch, sodiumcarboxylmethylcellulose solution, or suitable oil, the othercompositions can be prepared.

A lyophilized preparation may be prepared by dissolving activeingredients in a solution such as water, if necessary, with asolubilizer such as citric acid, edetic acid, polyphosphoric acid andtheir salts and a stabilizer such as mannitol, xylitol, sorbitol,glucose, fructose, lactose and maltose and lyophilizing it.

While a dosage differs dependent upon a state of disease, a route ofadministration, patient's age, and a body weight, it is usually 0.01 to50 mg/kg/day in case of oral administration in adult.

The method of the invention for inhibiting sPLA₂ mediated release offatty acids comprises contacting mammalian sPLA₂ with a therapeuticallyeffective amount of a pyrrolo[1,2-a]pyrazine sPLA₂ inhibitors (andformulation containing such inhibitors) as taught, supra.

Preferably compounds of the invention (per Formula (I) or (II) or (III)or (IV) or pharmaceutical formulations containing these compounds) arein unit dosage form for administration to a mammal. The unit dosage formcan be a capsule or tablet itself, or the appropriate number of any ofthese. The quantity of Active ingredient in a unit dose of compositionmay be varied or adjusted from about 0.1 to about 1000 milligrams ormore according to the particular treatment involved. It may beappreciated that it may be necessary to make routine variations to thedosage depending on the age and condition of the patient. The dosagewill also depend on the route of administration.

The improved method of treatment for sepsis using thepyrrolo[1,2-a]pyrazine sPLA₂ inhibitors (and formulation containing suchinhibitors) may be practiced as follows:

The inhibitors of this invention are given by injection, eithersubcutaneously or into muscle tissue or by injection into a vein.Intravenous injection is the preferred mode of delivery to the mammalbeing treated and offers the advantage of a quick effect and rapidaccess into the circulation system, particularly in emergencysituations.

It may be appreciated that it may be necessary to make routinevariations to the dosage depending on the age and condition of thepatient. The specific dose of a compound administered according to thisinvention to obtain therapeutic or prophylactic effects will, of course,be determined by the particular circumstances surrounding the case,including, for example, the compound administered, the route ofadministration and the condition being treated. Typical daily doses willcontain a non-toxic Compound (I) dosage level of from about 0.01 mg/kgto about 50 mg/kg of body weight of an Active ingredient of thisinvention.

This invention is a method of treating or preventing Inflammatorydiseased, (e.g., sepsis, rheumatoid arthritis, osteoarthritis, asthma)by administering to a mammal in need thereof a therapeutically effectiveamount inhibitor. The administration to a septic patient may be eithercontinuous or intermittent.

The decision to begin the therapy for sepsis will be based upon theappearance of the clinical manifestations of sepsis or laboratory testswhich show initiation of the sepsis cascade (inclusive of renalcomplications or coagulation abnormalities or multiple organ failure).Typical clinical manifestations are fever, chills, tachycardia,tachypnea, altered mental state, hypothermia, hyperthermia, acceleratedor repressed breathing or heart rates, increased or decreased whiteblood cell count, and hypotension. These and other symptoms are wellknown in the art as set out in standard references such as, Harrison'sPrinciples of Internal Medicine (ISBN 0-07-032370-4) 1994, pages511-515.

The decision to determine the length of therapy may be supported bystandard clinical laboratory results from commercially available assaysor instrumentation supporting the eradication of the symptoms definingsepsis. The method of the invention may be practiced by continuously orintermittently administering a therapeutically effective dose of theinhibitor. The administration can be conducted for up to a total ofabout 60 days with a preferred course of therapy lasting for up to 10days.

The decision to end therapy by the method of the invention may besupported by standard clinical laboratory results from commerciallyavailable assays or instrumentation or the disappearance of clinicalsymptoms characteristic of sepsis. The therapy may be restarted upon thereturn of sepsis. Pediatric forms of sepsis are also successfullytreated by the methods, compounds, and formulations of this invention.

When the compound of the present invention is a crystallized, it mayshow various crystal forms and crystal habits.

The present invention will be described in more detail in conjunctionwith examples and test examples hereinafter, but it is to be noted thatthe present invention is not limited thereto.

In the examples, the following abbreviations are used.

Me: methyl

Et: ethyl

Pr: propyl

Bu: butyl

Ph: phenyl

DBU: 1,8-diazabicyclo [5.4.0]-7-undecene

Boc: t-butyloxycarbonyl

DMSO: dimethylsulfoxide

BEST MODE FOR CARRYING OUT THE INVENTION Example 1

Example 1—Step 1

A mixture of 720 mg (5.81 mmol) of compound (1) and 904 mg (6.00 mmol)of 1-bromo-2-butanone was warmed at 60° C. for 20 hours to obtain aquaternary salt (compound (2)). NMR (CDCl₃) 67 1.17(t, J=7.2 Hz, 3H),2.77(s, 3H), 2.94(q, J=7.2 Hz, 2H), 4.19(s, 3H), 6.93(s, 2H), 8.57(d,J=3.9 Hz, 1H), 9.17(d, J=3.9 Hz, 1H).

Example 1—Step 2

To the crude compound (2) obtained in the step 1 were added 22 ml of1,2-dichloroethane and 1.32 g (8.72 mmol) of DBU, and the resultingmixture was heated at 70° C. and stirred in an oil bath for 20 hours. Tothe reaction solution were added chloroform, water, and brine toseparate an organic layer, and an aqueous layer was further extractedwith chloroform. The organic layer was combined, dried over magnesiumsulfate, thereafter the solvent is removed, and the residue wassubjected to silica gel column chromatography. The fractions elutingwith chloroform-methanol (100:1) were collected to give compound (3)(750 mg, 74% yield) as an oil.

NMR (CDCl₃) 67 1.27(t, J=7.4 Hz, 3H), 2.69(q, J=7.4 Hz, 2H), 4.04(s,3H), 6.62(s, 1H), 7.00-7.03(m, 1H), 7.16(d, J=0.8 Hz, 1H), 7.39-7.42(m,1H).

Example 1—Step 3

The compound (3) (2.4 g (13.6 mmol)) was dissolved in 15 ml of benzoylchloride, and 5.42 g (40.8 mmol) of aluminum chloride was added to thesolution at an internal temperature of −10° C. to 0° C. over 10 minutes.The resulting mixture was further stirred at 5° C. for 15 hours. Thereaction solution was poured into a mixed solution of ice-water andchloroform. The organic layer was separated and the aqueous layer wasextracted with chloroform. The organic layer was washed with aqueoussodium bicarbonate and brine, dried over magnesium sulfate, andconcentrated in vacuo and the residue was subjected to silica gelchromatography. The fractions eluting with chloroform-methanol (40:1)were collected to give compound (4) (2.68 g, 75% yield) as a crystal.The resulting crystal was recrystallized from ether and hexane. Meltingpoint: 83-84° C.

Elemental Analysis C₁₇H₁₆N₂O₂, Calcd.: C, 72.84; H, 5.75; N, 9.99 Found:C, 72.94; H, 5.78; N, 10.16

NMR (CDCl₃) 67 1.08(t, J=7.4 Hz, 3H), 2.33(q, J=7.4 Hz, 2H), 4.10(s,3H), 6.72(s, 1H), 7.31(s, 1H), 7.44-7.70(m, 5H), 8.66(d, J=0.9 Hz, 1H).IR (CHCl₃) 1615 cm⁻¹.

Example 1—Step 4

To a solution of 240 mg (1.8 mmol) of aluminum chloride in 12 ml ofmethylene chloride was added 312 mg (3.6 mmol) of boron-t-butylaminecomplex under ice-cooling over 3 minutes. The resulting mixture wasstirred under the same condition as described above for 10 minutes. Tothe resulting mixture was added dropwise a solution of 168 mg (0.6 mmol)of the compound (4) in 2.5 ml of methylene chloride under ice-cooling,thereafter the mixture was stirred for 20 minutes, and further stirredat room temperature for 3 hours. To the reaction mixture were addedchloroform, ice-water, and diluted hydrochloric acid, the admixture wasstirred for several minutes, thereafter the organic layer was separated,and the aqueous layer was further extracted with chloroform. The organiclayer was washed with brine, dried over magnesium sulfate, andconcentrated in vacuo. The residue was subjected to alumina columnchromatography. The fractions eluting, with chloroform-hexane (2:1) werecollected to give compound (5) (99 mg, 62% yield).

The compound was recrystallized from ether and hexane. Melting point:56-57° C.

Elemental Analysis C₁₇H₁₈N₂O, Calcd.: C, 76.66; H, 6.81; N, 10.52 Found:C, 76.47; H, 6.80; N, 10.53

NMR (CDCl₃) δ1.27(t, J=7.5 Hz, 3H), 2.69(q, J=7.5 Hz, 2H), 4.04(s, 3H),4.21(s, 2H), 6.73(s, 1H), 6.95-7.29(m, 7H).

Example 1—Step 5

To 1.7 g (6.38 mmol) of the compound (5) was added 51 ml of concentratedhydrochloric acid, and the resulting mixture was heated and stirred inan oil bath at 110° C. for 140 minutes. The reaction mixture wasconcentrated in vacuo. The residue was poured into a mixed solution ofice-water and chloroform, and sodium bicarbonate (12 g) was graduallyadded to the mixture. The organic layer was separated and the aqueouslayer was further extracted with chloroform. The organic layer waswashed with brine, dried over magnesium sulfate, and concentrated invacuo to obtain crude crystal of compound (6) (1.44 g, 89% yield). Thecrude crystal can be used for the next step. The crude crystal wasrecrystallized from ethyl acetate to give the compound of which meltingpoint is 204-207° C.

Elemental Analysis as C₁₇H₁₈N₂O₂, Calcd.: C, 72.32; H, 6.43; N, 9.92Found: C, 72.11; H, 6.48; N, 9.98

NMR (CDCl₃) δ1.24(t, J=7.6 Hz, 3H), 2.62(q, J=7.6 Hz, 2H), 4.16(s, 2H),6.35-6.41(m, 1H), 6.69-6.72(m, 1H), 7.01-7.29(m, 6H), 9.97(brs, 1H).

IR (CHCl₃) 3419, 3164, 1647 cm⁻¹.

Example 1—Step 6

To 1.18 g (4.68 mmol) of the compound (6) was added 35 ml of phosphorousoxychloride and the mixture was refluxed in oil bath for 4 hours. Theresidue obtained by distilling off excess phosphorus oxychloride wasdissolved in chloroform, and the mixture was poured into ice-water. Theresulting mixture was extracted with chloroform. The organic layer waswashed with water, dried over magnesium sulfate, and concentrated invacuo. The resulting residue was subjected to silica gel columnchromatography. The fractions eluting with chloroform-methanol (50:1)were collected to give compound (7) (1.15 g, 91% yield) as an oil.

NMR (CDCl₃) δ1.31(t, J=7.8 Hz, 3H), 2.75(q, J=7.8 Hz, 2H), 4.25(s, 2H),6.87(s, 1H), 6.99-7.02(m, 2H), 7.17-7.38(m, 5H).

Example 1—Step 7

To a suspension of methyl glycolate (2 ml) and sodium (200 mg (8.70mmol)) were added successively a solution of 250 mg (0.923 mmol) ofcompound (7) in 1 ml of methyl glycolate, and 25 mg of sodiump-toluenesulfinate, and the resulting mixture was heated at 90° C. inoil bath for 20 hours. The reaction mixture was diluted with chloroformand brine was added to the resulting mixture. The organic layer wasseparated and the aqueous layer was further extracted with chloroform.The organic layer was dried over magnesium sulfate and concentrated invacuo. The residue was subjected to silica gel column chromatography.The fractions eluting with ethyl acetate-hexane were collected to givecompound (8) (245 mg, 82% yield) as an oil.

NMR (CDCl₃) δ1.28(t, J=7.5 Hz, 3H), 2.71(q, J=7.5 Hz, 2H), 3.78(s, 3H),4.22(s, 2H), 5.01(s, 2H), 6.83(s, 1H), 6.89-7.29(m, 7H).

Example 1—Step 8

To a solution of 245 mg (0.756 mmol) of compound (8) in 11 ml of1,2-dichloroethane were added 480 mg (3.78 mmol) of oxalyl chloride and382 mg (3.78 mmol) of N-methylmorpholine, and the resulting mixture washeated at 50° C. in oil bath for 4 hours. The reaction mixture waspoured into aqueous ammonia, and the mixture was stirred at roomtemperature for 10 minutes, and then extracted with chloroform. Theorganic layer was washed with brine, dried over magnesium sulfate, andconcentrated in vacuo. The residue was subjected to alumina columnchromatography. The fractions eluting with chloroform were collected togive compound (I-1) (137 mg, 46% yield) as a crystal. The crude crystalwas recrystallized from a chloroform and methanol to obtain compound ofwhich melting point is 151-152° C.

Elemental Analysis C₂₁H₂₁N₃O₅, Calcd.: C, 63.79; H, 5.35; N, 10.63Found: C, 63.67; H, 5.56; N, 10.43

NMR (CDCl₃) δ1.21(t, J=7.5 Hz, 3H), 2.85(q, J=7.5 Hz, 2H), 3.75(s, 3H),4.24(s, 2H), 4.97(s, 2H), 5.70(brs, 1H), 6.68(brs, 1H), 7.06-7.14(m,3H), 7.23-7.31(m, 4H).

IR (CHCl₃) 3515, 3401, 1762, 1702, 1655 cm⁻¹.

Example 2

Example 2—Step 1

To a solution of 110 mg (0.278 mmol) of compound (I-1) in 15 ml ofmethanol was added 0.56 ml (0.556 mmol) of 1 N sodium hydroxide, and theresulting mixture was stirred at room temperature for 18 hours. Thereaction mixture was concentrated in vacuo, and ice-water was added tothe residue. To the resulting mixture was added 1 N hydrochloric acid(0.65 ml) and stirred at room temperature. The precipitated crystal wascollected by filtration to give compound (I-2) (90 mg, 85% yield). Theresulting crude crystal was recrystallized from methanol and chloroformto give compound of which decomposition point is 211-213° C.

NMR (DMSO-d6) δ1.07(t, J=7.2 Hz, 3H), 2.77(q, J=7.2 Hz, 2H), 4.34(s,2H), 4.65(s, 2H), 7.10-7.31(m, 6H), 7.46(brs, 1H), 7.73(d, J=4.8 Hz,1H), 8.03(brs, 1H)

IR (KBr) 3425, 1709, 1668, 1640 cm⁻¹.

The compounds (I-3) to (I-36) which were shown in Tables 1 to 4 weresynthesized in a manner similar to those described in Examples 1 and 2.

TABLE 1

Melting Com- point ¹H-NMR: δ pound No. R³⁶ R³⁷ R³⁸ (° C.) CDCl₃(R³⁶ =Me), DMSO-d₆(R³⁶ = H) I-3 Me cyclopropyl

177-179 3.75(s, 3H), 4.38(s, 2H), 4.96 (s, 2H), 7.09(d, J=4.8Hz, 1H),7.17(d, J=4.8Hz, 1H) I-4 H cyclopropyl

189-191 4.42(s, 2H), 4.79(s, 2H), 7.19 (d, J=5.1Hz, 1H), 7.71(d, J=5.1Hz, 1H) I-5 Me Et

165-166 3.76(s, 3H), 4.38(s, 2H), 4.97 (s, 2H), 7.19(d, J=4.8Hz, 1H),7.38(d, J=4.8Hz, 1H) I-6 H Et

225-227 4.55(s, 2H), 4.81(s, 2H), 7.29 (d, J=4.8Hz, 1H), 7.92(d, J=4.8Hz, 1H) I-7 Me Et

162-163 2.69(d, J=7.5Hz, 2H), 3.76(s, 3H), 4.96(s, 2H), 7.24(d, J=4.8Hz, 1H), 7.43(d, J=4.8Hz, 1H) I-8 H Et

205-206 2.78(d, J=7.0Hz, 2H), 4.80(s, 2H), 7.29(d, J=4.8Hz, 1H), 7.98(d,J=4.8Hz, 1H) I-9 Me Et

154-155 3.75(s, 3H), 3.76(s, 3H), 4.20 (s, 2H), 4.96(s, 2H), 7.13(d, J =4.8Hz, 1H), 7.26(d, J=4.8Hz, 1H) I-10 H Et

196-197.5 3.71(s, 3H), 4.32(s, 2H), 4.81 (s, 2H), 7.26(d, J=4.8Hz, 1H),7.80(d, J=4.8Hz, 1H)

TABLE 2

Com- Melting ¹H-NMR: δ pound No. R³⁶ R³⁷ R³⁸ point (° C.) CDCl₃(R³⁶ =Me), DMSO-d₆(R³⁶ = H) I-11 Me Et

144-148 3.75(s, 3H), 4.37(s, 2H), 4.97 (s, 2H), 7.11(d, J=5.1Hz, 1H),7.24(d, J=5.1Hz, 1H) I-12 H Et

209-211 4.49(s, 2H), 4.82(s, 2H), 7.25 (d, J=4.8Hz, 1H), 7.85(d, J=4.8Hz, 1H) I-13 Me Me

182-183 2.44(s, 3H), 3.75(s, 3H), 4.22 (s, 2H), 4.97(s, 2H) I-14 H Me

207-208 2.36(s, 3H), 4.34(s, 2H), 4.81 (s,2H) I-15 Me Et

165-166 3.76(s, 3H),2.82(d, J=7.5Hz, 2H), 4.96(s, 2H), 7.24(d, J= 4.8Hz,1H), 7.48(d, J=4.8Hz, 1H) I-16 H Et

203-205 2.91(d, J=7.8Hz, 2H), 4.80(s, 2H), 7.29(d, J=4.8Hz, 1H), 8.01(d,J=4.8Hz, 1H)

TABLE 3

Com- Melting ¹H-NMR: δ pound No. R³⁶ R³⁷ R³⁸ point (° C.) CDCl₃(R³⁶ =Me), DMSO-d₆(R³⁶ = H) I-17 Me Et

192-194 2.17(d, J=0.9Hz, 3H), 3.75(s, 3H), 4.20(s, 2H), 4.97(s, 2H) I-18H Et

207-208 2.18(s, 3H), 4.33(s, 2H), 4.81 (s, 2H), 7.71(s, 1H) I-19 Me Et

167-168 2.19(d, J=0.9Hz, 3H), 3.76(s, 3H), 4.17(s, 2H), 4.98(s, 2H) I-20H Et

204-205 2.18(d, J=0.9Hz, 3H), 4.31(s, 2H), 4.80(s, 2H), 7.72(d, J=0.9Hz, 1H) I-21 Me Et

164.5-165.5 2.12(d, J=0.9Hz, 3H), 3.73(s, 3H), 4.09(s, 2H), 4.94(s, 2H),7.72(d, J=0.9Hz, 1H) I-22 H Et

192-194 2.14(d, J=0.9Hz, 3H), 4.19(s, 2H), 4.80(s, 2H) I-23 Me Et

135-136.5 2.29(d, J=1.2Hz, 3H), 2.78(d, J=7.5Hz, 2H), 3.75(s, 3H),4.96(s, 2H), 7.28(d, J=1.2Hz, 1H) I-24 H Et

192-193 2.26(s, 3H), 2.87(d, J=7.5Hz, 2H), 4.80(s, 2H), 7.86(s, 1H) I-25Me Et

187-188 2.20(d, J=0.9Hz, 3H), 3.75(s, 3H), 4.20(s, 2H), 4.97(s, 2H) I-26H Et

218-219 2.20(s, 3H), 4.33(s, 2H), 4.81 (s, 2H), 7.77(s, 1H)

TABLE 4

Com- Melting ¹H-NMR: δ pound No. R³⁶ R³⁷ R³⁸ point (° C.) CDCl₃(R³⁶ =Me), DMSO-d₆(R³⁶ = H) I-27 Me Et

134.5-136 2.03(d, J=0.9Hz, 3H), 3.74(s, 3H), 4.03(s, 2H), 4.17(s, 2H),4.94(s, 2H), 6.39(d, J=0.9Hz, 1H) I-28 H Et

180.5-182.5 2.07(d, J=0.6Hz, 3H), 4.19(s, 2H), 4.21(s, 2H), 4.80(s, 2H),7.04(s, 1H) I-29 Me Et

147-149 2.13(d, J=0.9Hz, 3H), 3.74(s, 3H), 4.06(s, 2H), 4.95(s, 2H),6.75(d, J=0.9Hz, 1H) I-30 H Et

175-177 2.15(s, 3H), 4.18(s, 2H), 4.80 (s, 2H), 7.36(s, 1H) I-31 Me Et

161-163 2.21(d, J=0.9Hz, 3H), 3.76(s, 3H), 4.15(s, 2H), 4.98(s, 2H) I-32H Et

208-210 2.20(d, J=0.9Hz, 3H), 4.30(s, 2H), 4.81(s, 2H), 7.78(d, J=0.9Hz, 1H) I-33 Me Et

189-190 2.18(s, 3H), 3.75(s, 3H), 3.78 (s, 3H), 4.14(s, 2H), 4.97(s,2H), 7.06(s, 1H) I-34 H Et

200-201.5 2.18(d, J=0.6Hz, 3H), 3.69(s, 3H), 4.24(s, 2H), 4.80(s, 2H),7.06(s, 1H), 7.68(d, J=0.6Hz, 1H) I-35 Me Et

179.5-181 2.19(d, J=0.9Hz, 3H), 3.76(s, 3H), 4.20(s, 2H), 4.97(s, 2H),7.03(d, J=0.9Hz, 1H) I-36 H Et

190.5-193 2.19(d, J=0.9Hz, 3H), 4.35(s, 2H), 4.81(s, 2H), 7.74(d, J=0.9Hz, 1H)

Example 37

Example 37—Step 1

To a solution of 176 mg (0.430 mmol) compound (I-17) which wassynthesized from 2,5-dimethyl-3-methoxypyrazine (Heterocycles, 1992,34(9), 1759-1771) in accordance with the same manner as that of Example1 in 6 ml of 1,4-dioxane was added 100 mg (0.860 mmol) of seleniumdioxide, and the resulting mixture was refluxed or 9 hours. Theresulting mixture was concentrated in vacuo and the residue wassubjected to silica gel chromatography. The fractions eluting withchloroform-methanol (40:1) were collected to give compound (I-37) (63mg, 34% yield) as yellow crystal.

Melting Point: 201-202° C.

Elemental Analysis C₂₂H₂₃N₃O₆, Calcd.: C, 62.11; H, 5.45; N, 9.88 Found:C, 62.11; H, 5.46; N, 9.84

¹H-NMR (CDCl₃) δ1.20 (t, J=7.5 Hz, 3H), 2.17 (brs, 1H), 2.84 (q, J=7.5Hz, 2H), 3.75 (s, 3H), 4.23 (s, 2H), 4.43 (s, 2H), 4.97 (s, 2H),5.56(brs, 1H), 6.70 (brs, 1H), 7.03-7.10 (m, 2H), 7.20-7.33 (m, 4H).

IR (KBr) 3418, 3260, 1758, 1692, 1630, 1606, 1502, 1344, 1213, 1159cm⁻¹.

Example 38

Example 38—Step 1

To a solution of 19 mg of the compound (I-37) in 0.5 ml of methanol and0.5 ml of tetrahydrofuran was added 0.07 ml of 4 N sodium hydroxide atroom temperature, and the resulting mixture was stirred at the sametemperature for 1 hour. To the reaction mixture were added water and 1ml of 1 N hydrochloric acid, and the resulting mixture was extractedwith ethyl acetate. The organic layer was successively washed with waterand brine, dried over sodium sulfate, and the concentrated in vacuo. Theresidue was recrystallized from ethyl acetate and hexane to givecompound (I-38) (16 mg, 88% yield) as light yellow crystal.

Melting Point 211-212° C.

Elemental Analysis C₂₁H₂₁N₃O₆, Calcd.: C, 61.31; H, 5.14; N, 10.21Found: C, 61.16; H, 5.19; N, 10.13

¹H-NMR (DMSO-d₆) δ1.07 (t, J=7.5 Hz, 3H), 2.77(q, J=7.5 Hz, 2H), 4.31(d, J=3.0 Hz, 2H), 4.35 (s, 2H), 4.81 (s, 2H), 5.31 (brs, 1H), 7.10 (d,J=7.5 Hz, 2H), 7.21 (t, J=7.5 Hz, 1H), 7.30 (t, J=7.5 Hz, 2H), 7.50(brs, 1H), 7.68 (s, 1H), 7.89 (brs, 1H).

IR (KBr) 3412, 1712, 1667, 1501, 1317, 1227, 1212, 1163 cm⁻¹.

Example 39

Example 39—Step 1

L-valinol (9) (22.7 g (220 mmol)) was dissolved in 200 ml ofacetonitrile. To the mixture was added a solution of 41.7 g (220 mmol)of Boc-L-alanine in 100 ml of acetonitrile under ice-cooling.Thereafter, 46.6 g (242 mmol) of1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride was added,and the mixture was stirred at room temperature under nitrogenatmosphere for 24 hours. Acetonitrile was removed from the reactionmixture under reduced pressure, and the residue was poured into 100 mlof water, and extracted with ethyl acetate. The organic layer was washedwith brine, dried over magnesium sulfate, and concentrated in vacuo. Theresidue (77.5 g) was poured into diluted aqueous hydrochloric acid.After pH was adjusted to 2 to 3, the whole was extracted again withethyl acetate. The organic layer was washed with aqueous saturatedsodium bicarbonate, dried over magnesium sulfate, and concentrated invacuo to obtain compound (10) (35.7 g, 59% yield) as colorless crystal.A little quantities of the crystals were recrystallized from diethylether-hexane to obtain colorless needles.

Melting Point: 96.0-97.0° C.

Elemental Analysis as C₁₈H₂₆N₂O₄, Calcd.: C, 56.91; H, 9.55; N, 10.21Found: C, 56.77; H, 9.51; N, 10.14

¹H-NMR (CDCl₃) δ0.94 (dd, J=8.7, 6.9 Hz, 6H), 1.37 (d, J=7.2 Hz, 3H),1.45 (s, 9H), 1.89 (m, 1H), 2.28 (brs, 1H), 3.57-3.77 (m, 3H), 4.14(quint, J=7.2 Hz, 1H), 5.05 (d, J=6.6 Hz, 1H), 6.50 (d, J=7.5 Hz, 1H).

IR (CHCl₃) 3626, 3437, 1695, 1496, 1455, 1392, 1369, 1325 cm⁻¹.

Example 39—Step 2

The compound (10) (31.5 g (107 mmol)) was dissolved in 350 ml of ethylacetate. To the mixture were successively added 167 mg (1.07 mmol) ofTEMPO (2,2,6,6-tetramethylpiperidin-1-oxide), 1.27 g (10.7 mmol) ofpotassium bromide, and 268 ml of 0.4 N aqueous NaOCl (pH was adjusted to9.60 with NaHCO₃) at −6 ° C. (internal temperature) and the resultingmixture was stirred at the same temperature. After 45 minutes, thereaction mixture was poured into 100 ml of water, shaken, and extractedwith ethyl acetate. The ethyl acetate layer was washed with brine, andthe aqueous layer was further extracted with ethyl acetate. After dryingthe organic layers with magnesium sulfate, the solvent was removed underreduced pressure, and dried under reduced pressure to obtain 25.3 g oflight cream-colored foam. The residue was dissolved in 200 ml oftoluene, and the mixture was allowed to stand at room temperature for 2hours. The reaction mixture was concentrated under reduced pressure, andthe residue was subjected to silica gel chromatography. The fractionseluting with hexane-ethyl acetate (7:1) were collected to give compound(11) (14.1 g, 52% yield) as colorless crystal. A little quantities ofthe crystals were recrystallized from diethyl ether-hexane to obtaincolorless prisms.

Melting Point: 165.0-166.0° C.

Elemental Analysis C₁₃H₂₂N₂O₃, Calcd.: C, 61.39; H, 8.72; N, 11.01Found: C, 61.33; H, 8.74; N, 10.95

¹H-NMR (CDCl₃) δ1.15 (d, J=6.9 Hz, 6H), 1.27 (d, J=6.9 Hz, 3H), 1.50 (s,9H), 2.39 (m, 1H), 4.73 (m, 1H), 5.90 and 6.08 (each brs, total 1H),7.89 (brs, 1H).

IR (CHCl₃) 3408, 1685, 1472, 1454, 1437, 1395, 1370, 1325 cm⁻¹.

Example 39—Step 3

To a suspension of 1.02 g (3.99 mmol) of compound (11) in 5 ml of ethylacetate was added 10 ml (40.0 mmol) of 4 N hydrochloric acid in ethylacetate, and the resulting, mixture was stirred at room temperature.After 2 hours, the precipitated crystal was collected by filtration, andwashed with ethyl acetate to obtain compound (12) (655 mg, 86% yield) ascolorless crystal.

¹H-NMR (CD₃OD) δ1.57 (d, J=7.2 Hz, 3H), 1.77 (s, 6H), 3.92 (m, 1H), 4.13(q, J=7.2 Hz, 1H), 4.30 and 4.35 (each s, total 1H).

Example 39—Step 4

646 mg (3.39 mmol) of the compound (12) was dissolved in 2 ml of water,and sodium bicarbonate was gradually added to be alkaline. The reactionmixture was extracted with ethyl acetate, and further withdichloromethane. The organic layer was dried over magnesium sulfate,concentrated in vacuo, and dried under reduced pressure to obtain 517 mgof colorless crystal. The residue (517 g) was dissolved in 6.95 ml ofcyclohexene and 1.4 ml of methanol, and 290 mg of 10% Pd—C was added tothe mixture, and the resulting mixture was stirred at 80° C. for 3.5hours. After the reaction mixture was cooled to room temperature, Pd—Cwas filtered off. The filtrate was concentrated in vacuo. The residue(630 mg) was subjected to silica gel chromatography. The fractionseluting with toluene-ethyl acetate (1:1) were collected to give compound(13) (285 mg, 55% yield) as colorless crystal. A little quantities ofcrystals were recrystallized from diethyl ether-hexane to obtain ascolorless prisms.

Melting Point: 133.0-134.0° C.

Elemental Analysis C₈H₁₂N₂O.0.1 H₂O, Calcd.: C, 62.40; H, 7.99; N, 18.19Found: C, 62.61; H, 7.98; N, 18.24

¹H-NMR (CDCl₃) δ1.33 (d, J=7.2 Hz, 6H), 2.42 (s, 3H), 2.84 (m, 1H), 7.17(s, 1H), 12.48 (brs, 1H).

IR (CHCl₃) 3373, 1649, 1612, 1534, 1467, 1433, 1389, 1372 cm⁻¹.

Example 39—Steps 5 and 6

To 4.09 g (26.9 mmol) of the compound (13) was added 11.2 ml ofphosphorus oxychloride, and the mixture was refluxed under nitrogenatmosphere for 1 hour. After cooling the reaction mixture, the mixturewas gradually poured into 100 ml of ice-water and 60 ml of diethylether. To the mixture was added 45 ml of 28% aqueous ammonia to adjustpH to 5 to 6. About 40 ml of 5 N sodium hydroxide was further addedthereto to be alkaline, and then extracted with diethyl ether. Theorganic layer was dried over magnesium sulfate, and the solvent wasremoved under normal pressure to obtain 5.38 g of compound (14) as brownoil.

¹H-NMR (CDCl₃) δ1.32 (d, J=6.9 Hz, 6H), 2.62 (s, 3H), 3.06 (m, 1H), 8.26(s, 1H).

To a solution of 5.38 g compound (14) in 18 ml of methanol was added18.6 ml (93.0 mmol) of 28% sodium methoxide in methanol, and theresulting mixture was refluxed for 1 hour. After cooling the reactionmixture, it was concentrated in vacuo. The residue was poured into 30 mlof water, and the mixture was extracted with diethyl ether. The organiclayer was washed with brine and dried over magnesium sulfate. Thesolvent was removed under normal pressure to obtain compound (15) (4.27g, 96% yield) as brown oil.

¹H-NMR (CDCl₃) δ1.28 (d, J=6.9 Hz, 6H), 2.42 (s, 3H), 2.95 (m, 1H), 3.96(s, 3H), 7.85 (s, 1H).

Example 39—Step 7

Using the compound (15) as a starting material, compound (I-39) wassynthesized in a manner similar to that described in Example 1.

Example 40

The compound (I-40) was synthesized by carrying out the same reaction asdescribed in Example 2.

Example 41

Example 41—Step 1

Under ice-cooling, 7.24 g (84.0 mmol) of methacrylic acid, 16.3 g (84.0mmol) of 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride,and 7.54 g (84.0 mmol) of 2-amino-1-buthanol (16) were dissolved in 100ml of dichloromethane, and the mixture was stirred at room temperaturefor 20 hours. Dichloromethane was removed, then water was added to theresidue, and the whole was extracted with ethyl acetate. The organiclayer was washed successively with 10% hydrochloric acid, aqueoussaturated sodium bicarbonate, and brine, dried over sodium sulfate, andconcentrated in vacuo to obtain compound (17) (12.4 g, 93% yield) asyellow oil.

¹H-NMR (CDCl₃) δ0.98 (t, J=7.5 Hz, 3H), 1.59 (m, 2H), 1.98 (s, 3H),3.64(dd, J=11.1, 6.0 Hz, 1H), 3.74 (dd, J=11.1, 3.3 Hz, 1H), 3.93 (m,1H), 5.36 (s, 1H), 5.73 (s, 1H), 5.99 (brs, 1H).

IR (CHCl₃) 3428, 3004, 2962, 1711, 1655, 1617, 1514 cm⁻¹.

Example 41—Step 2

To a solution of 49.0 ml (690 mmol) of dimethyl sulfoxide in 50 ml ofdichloromethane was added gradually 29.5 ml (345 mmol) of oxalylchloride at −78° C. After stirring the mixture for 10 minutes, asolution of 18.1 g (115 mmol) of compound (17) in 100 ml ofdichloromethane was added, and the resulting mixture was stirred at −78°C. for 1 hour. To the mixture was added 96.0 ml (690 mmol) oftriethylamine, and the mixture was further stirred for 1 hour. To thereaction mixture was added 10% hydrochloric acid, and extracted withethyl acetate. The organic layer was dried over sodium sulfate andconcentrated in vacuo.

The residue was dissolved in 300 ml of dichloromethane and 100 ml ofmethanol, and ozone gas was bubbled through the solution at −78° C. Atthe time when a color of the reaction solution was turned to blue, 36.0ml (575 mmol) of dimethyl sulfide was added, and the resulting mixturewas stirred at room temperature for 2 hours. The reaction mixture waswashed with brine, dried over sodium sulfate, and concentrated in vacuo.

The residue was dissolved in 200 ml of ethanol. To the mixture was added17.7 g (230 mmol) of ammonium acetate and the resulting mixture wasrefluxed for 1 hour. After distilling off ethanol, water was added tothe residue, and extracted with ethyl acetate. The organic layer waswashed with brine, dried over sodium sulfate, and concentrated in vacuo.The residue was subjected to silica gel chromatography. The fractionseluting with ethyl acetate were collected to give compound (18) (3.19 g,20% yield) as white crystal.

Melting Point: 151.0-153.0° C.

FABMS (m/z) 139 ([M+H]⁺)

¹H-NMR (DMSO-d₆) δ1.14 (t, J=7.5 Hz, 3H), 2.41 (q, J=7.5 Hz, 2H), 2.21(s, 3H), 7.02 (s, 1H), 12.08 (brs, 1H).

IR (KBr) 2971, 2920, 1653, 1619, 1367 cm⁻¹.

Example 41—Step 3

A mixture of 2.72 g (19.7 mmol) of the compound (18) and 13.5 ml (145mmol) of phosphorus oxychloride was refluxed for 30 minutes. Thereaction mixture was gradually poured into ice-water, and neutralizedwith 4 N sodium hydroxide with stirring. The resulting mixture wasextracted with diethyl ether, and the organic layer was dried oversodium sulfate. To the residue obtained by distilling off the solventunder normal pressure was added 44.0 ml (44.0 mmol) of 1 N sodiummethoxide in methanol, and the mixture was refluxed for 5 hours.Methanol was removed under normal pressure, then water was added to theresidue, and extracted with diethyl ether. The organic layer was driedover sodium sulfate, and thereafter the solvent was removed under normalpressure to obtain compound (19) (1.32 g, 44% yield) as brown oil.

¹H-NMR (CDCl₃) δ1.29 (t, J=7.6 Hz, 3H), 2.42 (s, 3H), 2.69 (q, J=7.6 Hz,2H), 3.97 (s, 3H), 7.85 (s, 1H).

IR (CHCl₃) 2968, 1546, 1452, 1369 cm⁻¹.

Example 41—Step 4

Using the compound (19) as a starting material, compound (I-41) wassynthesized in a manner similar to that described in Example 1.

Compound (I-42) to compound (I-50) were synthesized by carrying out thesame reactions as described in Example 1 to Example 41. Results obtainedare shown in Tables 5 to 6.

TABLE 5

Com- Melting ¹H-NMR: δ pound No. R³⁶ R³⁸ R³⁹ point (° C.) CDCl₃(R³⁶ =Me), DMSO-d₆(R³⁶ = H) 1.39 Me

isopropyl 190-191 1.10(d, J=6.9Hz, 6H), 3.73 (s, 3H), 4.21(s, 2H),4.93(s, 2H) I-40 H

isopropyl 211-213 1.11(d, J=6.9Hz, 6H), 4.34 (s, 2H), 4.78(s, 2H),7.59(s, 1H) I-41 Me

Et 154-156 *3.65(s, 3H), 4.34(s, 2H), 4.87(s, 2H), 7.69(s, 1H) I-42 H

Et 197-198 4.34(s, 2H), 4.79(s, 2H), 7.67(s, 1H) I-43 Me

203-205 *3.68(s, 3H), 4.51(s, 2H), 4.99(s, 2H), 8.40(s, 1H) I-44 H

233-234 4.50(s, 2H), 4.91(s, 2H), 8.38(s, 1H) I-45 Me

isobutyl 129-130 2.25(d, J=7.2Hz, 2H), 3.72 (s, 3H), 4.22(s, 2H),4.94(s, 2H), 7.01(s, 1H) I-46 H

isobutyl 216-217 2.30(d, J=6.9Hz, 2H), 4.34 (s, 2H), 4.78(s, 2H),7.63(s, 1H) I-47 Me

Et 151-153 *3.65(s, 3H), 4.33(s, 2H), 4.87(s, 2H), 7.70(s, 1H) I-48 H

Et 202-204 4.33(s, 2H), 4.80(s, 2H), 7.68(s, 1H) *measured with DMSO-d₆

TABLE 6

Com- Melting ¹H-NMR: δ pound No. R³⁶ R³⁸ R³⁹ point (° C.) CDCl₃(R³⁶ =Me), DMSO-d₆(R³⁶ = H) I-49 Me

benzyl 178-180 *3.51(s, 3H), 3.76(s, 2H), 4.33(s, 2H), 4.82(s, 2H),7.84(s, 1H) I-50 H

benzyl 200-202 *3.77(s, 2H), 4.30(s, 2H), 4.78(s, 2H), 7.75(s, 1H)*measured with DMSO-d₆

Example 51

Example 51—Step 1

2-Thiopheneboronic acid (391 mg, 3.06 mmol) and 2 ml of 2M sodiumcarbonate were added to a solution of 800 mg of the compound (20) (2.04mmol) and 118 mg of tetrakis(triphenylphosphine) palladium (0.102 mmol)in 18 ml of dimethoxyethane—ethanol (5:1) under argon atmosphere, andthe resulting mixture was refluxed for 4 hours. To the reaction mixturewas added 12 ml of 1N hydrochloric acid, and the resulting mixture wasextracted with chloroform. The organic layer was washed with brine,dried over sodium sulfate, and concentrated in vacuo. The residue wassubjected to the silica gel column chromatography. The fractions elutingwith chloroform-methanol (98:2) were collected to obtain the compound(21) (592 mg, yield 83%) as a colorless crystal.

¹H-NMR(DMSO-d₆) δ1.04(t, J=7.5 Hz, 3H), 1.94(s, 3H), 2.36(q, J=7.5 Hz,2H), 4.20(s, 2H), 6.60(s, 1H), 6.65(m, 1H), 6.74(s, 1H), 7.19-7.68(m,6H), 10.40(brs, 1H).

Example 51—Step 2

Using the compound (21) as a starting material, compound (I-51) wassynthesized in a manner similar to that described in step 6 to step 8 ofExample 1.

Example 52

The compound (I-52) was synthesized by the same reaction described inExample 2 by using the compound (I-51) as a starting material.

Example 53

Example 53—Step 1

To a solution of 1 g of the compound (20) (2.55 mmol) in 10 ml ofdimethylformamide were added 339 mg of phenylacetylene (3.31 mmol), 59mg of dichlorobis(triphenylphosphine)palladium (0.084 mmol), 45 mg ofcooper (1) iode (0.24 mmol) and 490 mg of triethylamine (4.84 mmol). Theresulting mixture was stirred at 50° C. for 3 hours under argonatmosphere. After the reaction was completed, the resulting mixture wasadded to 2N hydrochloric acid and extracted with ethyl acetate. Theorganic layer was washed with brine, dried over sodium sulfate, andconcentrated in vacuo. The residue was purified by the silica gel columnchromatography to obtain the compound (22) (844 mg, yield 90%) as acolorless powder.

¹H-NMR(DMSO-d₆) δ1.08(3H, t, J=7.5 Hz), 1.93(3H, s), 2.46(2H, q, J=7.5Hz), 4.36(2H, s), 6.74(1H, m), 6.76(1H, s), 6.93(1H, s), 7.26-7.61(8H,m), 10.40(1H, br).

Example 53—Step 2

Using the compound (22) as a starting material, compound (I-53) wassynthesized in a manner similar to that described in step 6 to step 8 ofExample 1.

Example 54

The compound (I-54) was synthesized by the same reaction described inExample 2 by using the compound (I-53) as a starting material.

Example 55

Example 55—Step 1

Cooper (II) oxide (1.11 g, 14.0 mmol) was added to a solution of 1.47 gof the compound (20) (3.50 mmol), 490 mg of phenol (5.21 mmol) and 1.48g of potassium carbonate (10.5 mmol) in 7 ml of pyridine, and theresulting mixture was refluxed for 21 hours under nitrogen atmosphere.The reaction mixture was diluted with chloroform, filtered and thenremoved the solvent by distillation under reduced pressure. The residuewas diluted with ethyl acetate, washed 2 times with 1N sodiumhydrogensulfate, washed with brine, and then dried over sodium sulfate.The residue obtained by removing the solvent by distillation underreduced pressure was subjected to the silica gel column chromatography.The fractions eluting with n-hexane-ethyl acetate (5:1) were collectedto obtain the compound (23) (1.35 g, yield 100%) as a colorless oil.

¹H-NMR(CDCl₃) δ1.11(t, J=7.4 Hz, 3H), 2.28(q, J=7.4 Hz, 2H), 2.33(d,J=0.9 Hz, 3H), 4.08(s, 3H), 6.62(d, J=0.9 Hz, 1H), 6.79-7.05(m, 4H),7.17-7.24(m, 3H), 7.38-7.45(m, 2H), 8.75(s, 1H).

Example 55—Step 2

Using the compound (23) as a starting material, compound (I-55) wassynthesized in a manner similar to that described in step 6 to step 8 ofExample 1.

Example 56

The compound (I-56) was synthesized by the same reaction described inExample 2 by using the compound (I-55) as a starting material.

Example 57

Example 57—Step 1

The compound (24) (860 mg, 2.39 mmol) and 394 mg of phosphoruspentasulfide (2.77 mmol) were dissolved in 8 ml of pyridine, and theresulting mixture was refluxed for 3 hours. After the reaction wascompleted, the resulting mixture was diluted with water and extractedwith ethyl acetate. The organic layer was washed with 2N hydrochloricacid and brine successively, dried over sodium sulfate, and concentratedin vacuo. The residue was purified by the silica gel columnchromatography to obtain the compound (25) (559 mg, yield 62%) as ayellow crystal.

¹H-NMR(DMSO-d₆) δ1.02(3H, t, J=7.5 Hz), 2.06(3H, s), 2.33(2H, q, J=7.5Hz), 4.09(2H, s), 6.69(1H, d, J=7.5 Hz), 6.70(2H, s), 7.21-7.47(7H, m),12.02(1H, br).

Example 57—Step 2

To a solution of 250 mg of the compound (25) (0.66 mmol) in 5 ml ofdimethylformamide were added 275 mg of potassium carbonate (1.99 mmol),155 mg of t-butyl bromoacetate (0.79 mmol) and 11 mg of potassium iodide(0.066 mmol), and the resulting mixture was stirred at room temperaturefor 15 minutes. After the reaction was completed, the resulting mixturewas made acidic with 2N-hydrochloric acid and extracted with ethylacetate. The organic layer was dried over sodium sulfate, and thenremoved the solvent by distillation. The residue was purified by thesilica gel column chromatography to obtain the compound (26) (328 mg,yield 100%) as a yellow oil.

¹H-NMR(CDCl₃) δ1.20(3H, t, J=7.5 Hz), 1.46(9H, s), 2.19(3H, s), 2.57(2H,q, J=7.5 Hz), 3.97(2H, s), 4.04(2H, s), 6.63(1H, s), 6.69(1H, d, J=7.5Hz), 6.74(1H, s), 7.13-7.36(7H, m).

Example 57—Step 3

Using the compound (26) as a starting material, compound (I-57) wassynthesized in a manner similar to that described in step 8 of Example1.

¹H-NMR(CDCl₃) δ1.10(3H, t, J=7.5 Hz), 1.37(9H, s), 2.24(3H, d, J=0.9Hz), 2.70(2H, q, J=7.5 Hz), 3.89(2H, s), 4.07(2H, s), 5.67(1H, br),6.78(1H, d, J=7.5 Hz), 6.83(1H, d, J=0.9 Hz), 7.07(1H, br),7.15-7.38(7H, m).

Melting point: 138-139° C.

Example 58

Example 58—Step 1

The compound (I-57) (46 mg, 0.082 mmol) was dissolved in 3 ml ofdichloromethane. To the mixture was added 1 ml of trifluoroacetic acid,and the resulting mixture was stirred at room temperature for 4.5 hours.Trifluoroacetic acid was removed by distillation. To the residue wasadded water and the precipitated crystal was collected by filtration.The crystal was washed with water and dried to obtain the compound(I-58) (37 mg, yield 89%) as a yellow powder.

¹H-NMR(DMSO-d₆) δ0.86(3H, t, J=7.5 Hz), 2.25(3H, s), 2.50(2H, q, J=7.5Hz), 3.93(2H, s), 4.20(2H, s), 6.64(1H, d, J=6.6 Hz), 7.23-7.51(7H, m),7.48(1H, s), 7.82(1H, br), 8.20(1H, br).

Melting point: 103-105° C.

Example 59

Example 59—Steps 1 to 3

To a solution of 1.01 g of the compound (27) (4.94 mmol) in 20 ml oftetrahydrofuran was added dropwise 3.90 ml of n-butyllithium in hexane(1.53 M, 5.97 mmol) at −20° C., and the resulting mixture was stirredfor 30 minutes in the same condition. To the mixture was added 0.795 mlof 4-fluorobenzaldehyde (7.41 mmol) at −20° C. and the resulting mixturewas stirred for 15 minutes in the same condition. To the reactionmixture were added 5 ml of aqueous ammonium chloride, 5 ml of water andethyl acetate under ice-cooling. The organic layer was separated and theaqueous layer was extracted with ethyl acetate. The organic layer waswashed with water and brine and dried over sodium sulfate. The oilyresidue (the compound (28)) obtained by removing the solvent bydistillation under reduced pressure was subjected to the next reactionwithout any purification.

Chlorotrimethylsilane (7.95 ml, 62.6 mmol) was added slowly to asuspension of 9.44 g of sodium iodide (63.0 mmol) in 11 ml ofacetonitrile at room temperature, and the resulting mixture was stirredfor 15 minutes in the same condition. To the mixture was added slowly asolution of the compound (28) obtained above step in 15 ml ofacetonitrile under ice-cooling, and the resulting mixture was stirred atroom temperature for 2.75 hours. The reaction mixture was poured into amixture of ice water and ethyl acetate to separate the organic layer.The aqueous layer was extracted with ethyl acetate. The organic layerwas successively washed with 25 ml of aqueous sodium hydrogencarbonate,25 ml of 10% sodium thiosulfate and 25 ml of brine, and dried oversodium sulfate. The oily residue (the compound (29)) obtained byremoving the solvent by distillation under reduced pressure wassubjected to next reaction without any purification.

To the compound (29) obtained as described above was added 15 ml of 36%hydrochloric acid at room temperature, and the resulting mixture wasrefluxed for 30 minutes. To the mixture was added 15 ml of water underice-cooling. The insoluble substance was collected by filtration, washedwith water, ether, and then dried under reduced pressure to obtain thecompound (30) (1.08 g, yield 81% as a colorless powder.

¹H-NMR(CDCl₃) δ2.22(s, 3H), 2.44(s, 3H), 4.12(s, 2H), 7.02-7.16(m, 7H).

Example 59—Steps 4 to 5

Phosphorus oxychloride (2 ml) was added to 1.00 g of the compound (30)(3.70 mmol) at room temperature, the resulting mixture was refluxed for15 minutes, and then excess phosphorus oxychloride was removed bydistillation under reduced pressure. Ice was added to the residue, andthe mixture was extracted with ethyl acetate. The organic layer waswashed 2 times with 10 ml of aqueous sodium hydrogencarbonate, with 10ml of water, and 10 ml of brine, and then dried over sodium sulfate. Thecrystalline residue (the compound (31)) obtained by removing the solventby distillation under reduced pressure was subjected to next reactionwithout any purification.

To a suspension of the compound (31) and 1.32 g of sodiump-toluenesulfinate (7.41 mmol) in 10 ml of ethanol was added 0.11 ml of1N hydrochloric acid (0.11 mmol) at room temperature, and the resultingmixture was refluxed for 6 hours. The reaction mixture was cooled underice-cooling. The precipitated crystal was collected by filtration,washed 4 times with 2.5 ml of cold ethanol, and then dried under reducedpressure to obtain the compound (32) (1.28 g, yield 85%).

¹H-NMR(CDCl₃) δ2.31(s, 3H), 2.41(s, 3H), 2.54(s, 3H), 4.22(s, 2H),6.92-7.07(m, 3H), 7.30-7.35(m, 3H), 8.02(d, J=8.4 Hz, 2H).

Example 59—Step 6

Methyl glycolate (0.675 ml, 8.57 mmol) was added slowly to a suspensionof 249 mg of sodium hydride (60%, 6.21 mmol) in 10 ml ofdimethylformamide under ice-cooling, and the resulting mixture wasstirred at room temperature for 10 minutes. To the resulting mixture wasadded 1.00 g of the compound (32) (2.45 mmol) at room temperature, andthe resulting mixture was stirred for 50 minutes in the same condition.The reaction mixture was poured into a mixture of 10% hydrochloric acid,ice water and ether to separate the organic layer. The aqueous layer wasextracted with ether. The organic layer was washed successively with 20ml of aqueous sodium hydrogencarbonate, 20 ml of water, and 20 ml ofbrine, and then dried over sodium sulfate. Hexane was added to thecrystalline residue obtained by removing the solvent by distillationunder reduced pressure, and the mixture was allowed to warm to produceslurry. The crystal was collected by filtration, washed with hexane, andthen dried under reduced pressure to obtain the compound (33) (654 mg,,yield 78%).

¹H-NMR(CDCl₃) δ2.22(s, 3H), 2.35(s, 3H), 3.79(s, 3H), 4.14(s, 2H),5.03(s, 2H), 6.65(dd, J=0.8, 1.4 Hz, 1H), 6.87(dd, J=0.8, 1.4 Hz, 1H),6.96(m, 2H), 7.09(m, 2 H).

Example 59—Step 7

Oxalyl chloride (0.460 ml, 5.27 mmol) was added dropwise to a solutionof 565 mg of the compound (33) (1.65 mmol) and 0.580 ml ofN-methylmorpholine (5.28 mmol) in 5.5 ml of methylene chloride underice-cooling, and the resulting mixture was stirred for 30 minutes in thesame condition. The reaction mixture was poured into a mixture of 2 mlof 28% aqueous ammonia, 5 ml of ice water and ethyl acetate. Theinsoluble substance was removed by Celite filtration. To the filtrationwas added 8 ml of 10% aqueous hydrochloric acid. The organic layer wasseparated, washed with water and brine, dried over sodium sulfate, andconcentrated in vacuo. The residue was subjected to the silica gelcolumn chromatography. The fractions eluting with ethyl acetate werecollected to obtain the compound (I-59) (35.4 mg, yield 5%) as acrystal. The resulting crystal was recrystallized from ethyl acetate andhexane. Melting point : 212-214° C.

¹H-NMR(CDCl₃) δ2.30(d, J=0.9 Hz, 3H), 2.40(s, 3H), 3.77(s, 3H), 4.18(s,2H), 5.00(s, 2H), 5.50(brs, 1H), 6.60(brs, 1H), 6.92(d, J=0.9 Hz, 1H),6.95-7.11(m, 4H).

Example 60

Example 60—Step 1

4N sodium hydroxide (0.0500 ml, 0.200 mmol) was added to a mixture of19.8 mg of the compound (I-59) (0.0479 mmol), 0.5 ml of methanol and 0.5ml of tetrahydrofuran at room temperature, and the resulting mixture wasstirred for 30 minutes in the same condition. To the mixture was added0.5 ml of 1N hydrochloric acid under ice-cooling, and the resultingmixture was extracted with ethyl acetate. The organic layer was washedwith water and brine, dried over sodium sulfate, and concentrated invacuo. The residue was recrystallized from ethyl acetate, methanol ,andhexane to obtain the compound (I-60) (19.0 mg, yield 99%) as a crystal.Melting point: 239.5-242.5° C.

¹H-NMR(DMSO-d₆) δ2.24(s, 3H), 2.38(s, 3H), 4.33(s, 2H), 4.82(s, 2H),7.12(m, 2H), 7.24(m, 2H), 7.46(d, J=0.9 Hz, 1H), 7.48(brs, 1H),7.85(brs, 1H).

Example 61

Example 61—Step 1

The compound (34) (18.2 g, 0.160 mol) and 9.43 g of 90% acetaldehyde(0.190 mol) were dissolved in 20 ml of acetic acid. To the resultingmixture was added a mixture of 300 mg of 10% palladium-carbon catalystand 0.63 ml of piperidine (6.37 mmol) in 10 ml of acetic acid, and themixture was stirred at room temperature for 3 hours with retaining 1 to2 atm of pressure under hydrogen atmosphere. The catalyst was filteredoff. The filtration was diluted with toluene, washed with water, andthen distilled under reduced pressure to obtain the compound (35) (20.0g, yield 88%) showing the boiling point of 92-94° C. (13 mmHg) as acolorless oil (refer to OS, III, 385, 1955; J. Am. Chem. Soc., 66, 8861944)).

Example 61—Step 2

A mixture of 46.0 g of the compound (35) (0.326 mol), 77.1 g of bromoacetaldehyde diethylacetal (0.391 mol), 54.0 g of potassium carbonate(0.391 mol) and dimethylformamide (230 ml) was stirred with heating at70° C. for 72 hours under nitrogen atmosphere. Dimethylformamide wasremoved by distillation under reduced pressure. Water was added to theresidue, the mixture was extracted with toluene. The organic layer waswashed with water, dried over magnesium sulfate, and the solvent wasremoved. The residue was distilled under reduced pressure to obtain thecompound (36) showing boiling point of 105-106° C. (1 mmHg) (44.3 g,yield 56%) as a colorless oil.

¹H-NMR(CDCl₃) δ1.07(t, J=7.4 Hz, 3H), 1.18(t, J=7.0 Hz, 3H), 1.21(t,J=7.0 Hz, 3H), 1.33(t, J=7.0 Hz, 3H), 1.74-2.08(m, 3H), 2.39(dd, J=13.6,8.2 Hz, 1H), 3.45-3.76(m, 4H), 4.16-4.33(m, 2H), 4.77(dd, J=8.2, 4.0 Hz,1H).

Example 61—Step 3

A mixture of 168.2 g of the compound (36) (0.691 mol), 74.6 g ofpotassium acetate (0.760 mol) and dimethyl sulfoxide (336 ml) was heatedunder nitrogen atmosphere in an oil bath at 160° C. for 15 hours. Aftercooling, water was added, and the mixture was extracted with ether. Theorganic layer was washed with water, dried over magnesium sulfate, andthe solvent was removed. The residue was distilled under reducedpressure to obtain the compound (37) showing boiling point of 133-137°C. (33 mmHg) as a colorless oil (112.4 g, yield 88%).

¹H-NMR(CDCl₃) δ1.09(t, J=7.0 Hz, 3H), 1.22(t, J=7.0 Hz, 3H), 1.23(t,J=7.0 Hz, 3H), 1.58-1.99(m, 4H), 2.59-2.73(m, 1H), 3.48-3.81(m, 4H),4.68(dd, J=7.4, 4.2 Hz, 1H).

Example 61—Step 4

To a suspension of 1.53 g of magnesium (63.0 mmol) and 0.26 ml of1,2-dibromoethane (3.00 mmol) in 50 ml of ether was added dropwise asolution of 12.2 g of 2-biphenylmethyl chloride (60.0 mmol) in 24 ml ofether under ice-cooling. The resulting mixture was allowed to warm toroom temperature, and stirred until magnesium was dissolved. A solutionof 9.26 g of the compound (37) (50.0 mmol) in 28 ml of ether was addedto the mixture at room temperature, the resulting mixture was stirredfor 16 hours and then refluxed for 3 hours. Aqueous solution (25 mnl) ofammonium chloride (5.35 g) as added to the reaction mixture underice-cooling, the resulting mixture was made acidic with 63 ml of 2Nsulfuric acid, and stirred under ice-cooling for 30 minutes, further atroom temperature for 30 minutes. The reaction mixture was neutralized bysodium hydrogencarbonate, and extracted with toluene. The organic layerwas washed with brine, dried over magnesium sulfate, and concentrated invacuo. The residue was subjected to the silica gel chromatography. Thefractions eluting with ethyl acetate: toluene (1:9) were collected toobtain the compound (38) as a colorless oil (17.6 g, yield 99%).

¹H-NMR(CDCl₃) δ0.68(t, J=7.2 Hz, 3H), 1.12(t, J=6.9 Hz, 3H), 1.15(t,J=7.2 Hz, 3H), 1.21-1.44(m, 2H), 1.50-1.62(m, 1H), 1.87-1.96(m, 1H),2.50(m, 1H), 3.24 —3.58(m, 4H), 3.74(d, J=16.8Hz, 1H), 3.82(d, J=16.8Hz,1H), 4.27(t, J=6.0 Hz, 1H), 7.15—7.42(m, 9H).

Example 61—Step 5

To a solution of 3.00 g of the compound (38) (8.50 mmol) in 30 ml oftetrahydrofuran was added 5 ml of 2N hydrochloric acid at roomtemperature, and the resulting mixture was stirred at the sametemperature for 3 hours. The reaction mixture was poured into water, themixture was extracted with ether, and the organic layer was washed withwater, dried, and concentrated in vacuo. The residue was dissolved in 30ml of tetrahydrofuran. To the resulting mixture was added allylamine(0.77 ml, 10.2 mmol) under ice-cooling, and the mixture was stirred atthe same temperature for 1 hour. After evaporation to dryness underreduced pressure, the residue was subjected to the silica gelchromatography. The fractions eluting with hexane-hexane/ethyl acetate(50/1) were collected to obtain the compound (39) (1.92 g, yield 75%) asa colorless oil.

¹H-NMR (300M, CDCl₃): 1.15 (3H, t, J=7.8 Hz), 2.42 (2H, q, J=7.8 Hz),3.82(2H, s), 4.00 (2H, d, J=6.0 Hz), 4.73 (1H, d, J=17.5 Hz), 4.91 (1H,d, J=10.2 Hz), 5.53 (1H, m), 6.05 (1H, s), 6.51 (1H, s), 6.87 (1H, m),7.20-7.50 (8H, m).

Example 61—Step 6

To a solution of 200 mg of the compound (39) (0.67 mmol) in 2 ml oftoluene were added 0.104 ml of methyl chlorocarbonate (1.34 mmol) and153 mg of aluminum chloride (1.00 mmol) at room temperature, and themixture was stirred at the same temperature for 30 minutes. The reactionmixture was. poured into water, extracted with ether, and the organiclayer was washed with water, dried, and concentrated in vacuo. Theresidue was subjected to the silica gel column chromatography. Thefractions eluting with hexane/ethyl acetate (10/1) were collected toobtain the compound (40) (140 mg, yield 59%) as a colorless oil.

¹H-NMR (300M, CDCl₃): 1.14 (3H, t, J=7.8 Hz), 2.38 (2H, q, J=7.8 Hz),3.76 (3H, s), 3.83 (2H, s), 4.46 (1H, d, J=17.1 Hz), 4.60 (2H, m), 4.83(1H, d, J=10.5 Hz), 5.64(1H, m), 6.82 (1H, d, J=8.1 Hz), 6.89 (1H, s),7.20-7.50 ( 8H, s).

Example 61—Step 7

To a solution of 710 mg of the compound (40) (1.98 mmol) in 7 ml ofacetonitrile was added 1.00 g of iodine (7.92 mmol) at room temperature,and the mixture was stirred at the same temperature for 20 hours. Ethylacetate was poured into the reaction mixture, and the resulting mixturewas washed with aqueous sodium sulfite, further with water, dried, andconcentrated in vacuo. The residue was dissolved in hexane/ethyl acetate(1/1), and passed through the silica gel layer. The eluent wasconcentrated in vacuo to obtain the compound (41) (919 mg, yield 99%) asa colorless amorphous.

¹H-NMR (300M, CDCl₃): 1.13 (3H, t, J=7.5 Hz), 2.40 ( 2H, t, J=7.5 Hz),3.15 (1H, t, J=7.59 Hz), 3.40 (2H, m), 3.80 ( 1H, m), 3.87 (1H, d,J=17.1 Hz), 3.92 (1H, d, J=17.1 Hz), 4.44 ( 1H, m), 6.87 ( 1H, m), 6.98( 1H, s), 7.20-7.50 ( 8H, m).

Example 61—Step 8

To a solution of 900 mg of the compound (41) (1.91 mmol) in 10 ml oftoluene was added 0.43 ml of 1,8-diazabicyclo[5.4.0]-7-undecene (2.88mmol) at room temperature, and the mixture was stirred at 80° C. for 1hour. The solvent was removed by distillation and the residue wassubjected to the silica gel chromatography. The fractions eluting withhexane/ethyl acetate (4/1)—(2/1) to obtain the compound (42) (620 mg,yield 95%) as a colorless oil.

¹H-NMR (CDCl₃) 1.12(3H, t, J=7.5 Hz), 2.37(2H, q, J=7.5 Hz), 3.91( 2H,s), 4.06(2H, s), 4.41(1H, d, J=2.1 Hz), 4.87(1H, d, J=2.1 Hz), 6.88(1H,d, J=7.5 Hz), 7.00 (1H, s), 7.30-7.50(8H, m).

Example 61—Step 9

To a solution of 550 mg of the compound (42) (1.61 mmol) in 10 ml of 99%ethanol was added 3.72 g of ammonium acetate, and the mixture wasrefluxed for 20 hours. The mixture was concentrated in vacuo. Theresidue was washed with water, dissolved in chloroform. Further, ethylacetate was added to the mixture and concentrated. The precipitatedcrystal was collected by filtration to obtain the compound (43) (338 mg,yield 62%) as a colorless crystal. Melting point: 238-239° C.

¹H-NMR(DMSO-d₆) 1.02(3H, t, J=7.5Hz), 1.93(3H, s), 2.33(2H, q, J=7.5Hz),4.03(2H, s), 6.50 (1H, s), 6.69 (1H, d, J=6.6Hz), 6.70 (1H, s),7.20-7.50 (8H, m), 10.35 (1H, s).

Example 61—Step 10

Using the compound (43) as a starting material, compound (I-21) wassynthesized in a manner similar to that described in step 6 to step 8 ofExample 1.

Example 62

The compound (I-22) was synthesized by the same reaction described inExample 2 by using the compound (I-21) as a starting material.

Example 63

Example 63—Step 1

To a solution of 7.65 g of aluminum chloride (57.4 mmol) in 60 ml ofnitromethane was added dropwise 6.65 ml of benzoyl chloride (57.3 mmol)under ice-cooling, and the mixture was stirred for 15 minutes in thesame condition. To the mixture was added dropwise a solution of 2.93 gof the compound (44) (which can be synthesized in accordance with themethod described in Our. J. Med. Chem., 28, 481 (1993)) in 40 ml ofnitromethane under ice-cooling over 20 minutes, and the resultingmixture was stirred for 30 minutes in the same condition, furtherstirred at room temperature for 30 minutes. The reaction mixture waspoured into a mixture of ice water and ethyl acetate to separate theorganic layer. The aqueous layer was extracted with ethyl acetate. Theorganic layer was washed with 10 ml of 28% aqueous ammonia, 2 times withwater, and with brine, dried over sodium sulfate, and concentrated invacuo. The residue was subjected to the silica gel columnchromatography. The fractions eluting with n-hexane-ethyl acetate (4:1)were collected to obtain the compound (45) (4.20 g, yield 85%,) as acolorless oil.

¹H-NMR(CDCl₃) δ1.14(t, J=7.5 Hz, 3H), 2.55(qd, J=7.5, 0.6 Hz, 2H),3.89(s, 3H), 6.85(dt, 1H, J=2.7, 0.6 Hz), 7.46-7.53(m, 2H), 7.59(m, 1H),7.71(m, 2H), 9.48 (brs, 1H).

Example 63—Step 2

To a solution of 776 mg of the compound (45) (3.02 mmol) in 15 ml ofmethanol was added 134 mg of sodium borohydride (3.55 mmol) underice-cooling, and the mixture was stirred for 20 minutes in the samecondition. Aqueous ammonium chloride (3 ml), water and ethyl acetatewere added to the reaction mixture under ice-cooling to separate theorganic layer. The aqueous layer was extracted with ethyl acetate. Theorganic layer was washed with water, brine, dried over sodium sulfate,and concentrated in vacuo. The residue was subjected to the nextreaction without any purification.

To a suspension of 2.70 g of sodium iodide (18.0 mmol) in 3 ml ofacetonitrile was added slowly 2.30 ml of chlorotrimethylsilane (18.1mmol) at room temperature, and the mixture was stirred for 15 minutes inthe same condition. To the mixture was added slowly a solution of theresidue obtained above in 9 ml of acetonitrile under ice-cooling, andthe resulting mixture was stirred at room temperature for 35 minutes. INsodium hydroxide (10.5 ml) was added to the reaction mixture underice-cooling, and resulting mixture was extracted 2 times with 30 ml ofethyl acetate. The organic layer was washed successively with 30 ml of3% aqueous sodium thiosulfate, 30 ml of water, and 15 ml of brine, driedover sodium sulfate, and concentrated in vacuo. The residue wassubjected to silica gel column chromatography. The fractions elutingwith n-hexane-ethyl acetate (5:1) were collected to obtain the compound(46) (647 mg, yield 88%) as a colorless crystal.

¹H-NMR(CDCl₃) δ1.17(t, J=7.5 Hz, 3H), 2.45(q, J=7.5, 2H), 3.78(s, 3H),3.94(s, 2H), 6.78(d, 1H, J=2.7 Hz), 7.12-7.17(m, 2H), 7.20-7.34(m, 3H),8.56(brs, 1H).

Example 63—Step 3

A solution of 104 mg of the compound (46) (0.427 mmol) in 2 ml ofdimethylformamide was added dropwise to 26.2 mg of sodium hydride (60%)(0.655 mmol) under ice-cooling, and the mixture was stirred at roomtemperature for 30 minutes. To the resulting mixture was added 0.0554 mlof allyl bromide (0.640 mmol) in the same condition, and stirred for 1hour. Water and ether were added into the reaction mixture underice-cooling to separate the organic layer. The aqueous layer wasextracted with ether. The organic layer was washed with water and brine,dried over sodium sulfate, and concentrated in vacuo. The residue wassubjected to the silica gel chromatography. The fractions eluting withn-hexane-ethyl acetate (10:1) were collected to obtain the compound (47)(80.4 mg, yield 66%) as a colorless oil. ¹H-NMR(CDCl₃) δ1.16(t. J=7.5Hz. 3H), 2.44(q, J=7.5 Hz,. 2H), 3.78(s,. 3H). 3.95(s, 2H), 4.70(ddt,J=17.1. 1.6, 1.6 Hz. 1H), 4.80(dt, J=4.8. 1.6 Hz, 2H). 5.01(ddt, J=10.2,1.6 1.6 Hz, 1H), 5.84(ddt, J=17.1. 10.2, 4.8 Hz, 1H). 6.93(s, 1H),7.02(m. 2H). 7.13-7.30(tm L3H).

Example 63—Step 4

The compound (I-17) was synthesized by the same reaction described in 7to 10 step of Example 61 by using the compound (47) as a startingmaterial.

Example 64

The compound (I-18) was synthesized by the same reaction described inExample 2 by using the compound (I-17) as a starting material.

The compounds (I-61) to (I-106) were synthesized by the same reactiondescribed in Examples 1 to 64. The results are shown in Tables 7 to 13.

TABLE 7

Com- Melting ¹H-NMR: δ pound No. R³⁶ R³⁸ point (° C.) CDCl₃(R³⁶ = Me),DMSO-d₆(R³⁶ = H) I-51 Me

134.5-136 2.03(d, J=0.9Hz, 3H), 3.74(s, 3H), 4.03(s, 2H), 4.17(s, 2H),4.94(s, 2H), 6.39(d, J=0.9Hz, 1H) I-52 H

180.5-182.5 2.07(d, J=0.6Hz, 3H), 4.19(s, 2H), 4.21(s, 2H), 4.80(s, 2H),7.04(s, 1H) I-53 Me

147-149 2.13(d, J=0.9Hz, 3H), 3.74(s, 3H), 4.06(s, 2H), 4.95(s, 2H),6.75(d, J=0.9Hz, 1H) I-54 H

175-177 2.15(s, 3H), 4.18(s, 2H), 4.80 (s, 2H), 7.36(s, 1H) I-55 Me

161-163 2.21(d, J=0.9Hz, 3H), 3.76(s, 3H), 4.15(s, 2H), 4.98(s, 2H) I-56H

208-210 2.20(d, J=0.9Hz, 3H), 4.30(s, 2H), 4.81(s, 2H), 7.78(d, J=0.9Hz, 1H) I-61 Me

189-190 2.18(s, 3H), 3.75(s, 3H), 3.78 (s, 3H), 4.14(s, 2H), 4.97(s,2H), 7.06(s, 1H) I-62 H

200-201.5 2.18(d, J=0.6Hz, 3H), 3.69(s, 3H), 4.24(s, 2H), 4.80(s, 2H),7.06(s, 1H), 7.68(d, J=0.6Hz, 1H) I-63 Me

179.5-181 2.19(d, J=0.9Hz, 3H), 3.76(s, 3H), 4.20(s, 2H), 4.97(s, 2H),7.03(d, J=0.9Hz, 1H) I-64 H

190.5-193 2.19(d, J=0.9Hz, 3H), 4.35(s, 2H), 4.81(s, 2H), 7.74(d, J=0.9Hz, 1H) *measured with DMSO-d₆

TABLE 8

Com- Melting ¹H-NMR: δ pound No. R³⁶ R³⁸ point (° C.) CDCl₃(R³⁶ = Me),DMSO-d₆(R³⁶ = H) I-65 Me

*2.14(d, J=0.6Hz, 3H), 3.67(s, 3H), 4.22(s, 2H), 4.88(s, 2H), 7.31(d,J=0.6Hz, 1H) I-66 H

2.15(s, 3H), 4.21(s, 2H), 4.72 (s, 2H), 7.38(s, 1H) I-67 Me

2.12(d, J=1.2Hz, 3H), 3.74(s, 3H), 3.88(s, 3H), 4.09(s, 2H), 4.94(s,2H), 6.76(d, J=12Hz, 1H) I-68 H

2.14(s, 3H), 3.80(s, 3H), 4.20 (s, 2H), 4.79(s, 2H), 7.32(s, 1H) I-69 Me

167.5-169.5 2.17(d, J=0.9Hz, 3H), 2.44(s, 3H), 3.74(s, 3H), 4.09(s, 2H),4.94(s, 2H), 6.76(d, J=0.9Hz, 1H) I-70 H

179.5-181.5 2.14(s, 3H), 2.36(s, 3H), 4.19 (s, 2H), 4.79(s, 2H), 7.34(s,1H) I-71 Me

190-192 2.16(d, J=0.6Hz, 3H), 3.75(s, 3H), 4.27(s, 2H), 4.95(s, 2H),7.20(d, J=0.6Hz, 1H) I-72 H

131-133 2.14(s, 3H), 4.32(s, 2H), 4.79 (s, 2H), 7.39(s, 1H) I-73 Me

215-217 *2.14(d, J=0.9Hz, 3H), 3.67(s, 3H), 4.22(s, 2H), 4.90(s, 2H),7.44(d, J=0.9Hz, 1H) I-74 H

189-191 2.15(s, 3H), 4.21(s, 2H), 4.82 (s, 2H), 7.40(s, 1H) *measuredwith DMSO-d₆

TABLE 9

Com- Melting ¹H-NMR: δ pound No. R³⁶ R³⁷ R³⁸ R³⁹ point (° C.) CDCl₃(R³⁶= Me), DMSO-d₆(R³⁶ = H) I-75 Me cyclo- propyl

Me 210-211.5 2.15(d, J=0.9Hz, 3H), 3.74(s, 3H), 4.34(s, 2H), 4.96(s,2H), 6.99 (d, J=0.9Hz, 1H) I-76 H cyclo- propyl

Me 194.5-196 2.15(d, J=0.6Hz, 3H), 4.38(s, 2H), 4.79(s, 2H), 7.62(d,J=0.6Hz, 1H) I-77 Me cyclo- propyl

Me 179-182.5 2.16(1, J=0.9Hz, 3H), 3.75(s, 3H), 4.31(s, 2H), (s, 2H),6.96 (d, J=0.9Hz, 1H) I-78 H cyclo- propyl

Me 185-187 2.16(d, J=0.9Hz, 3H), 4.37(s, 2H), 4.79(s, 2H), 7.64(d,J=0.9Hz, 1H) I-79 Me Et

193-194 3.74(s, 3H), 4.21(s, 2H), 4.93(s, 2H), 7.05 (s, 1H) I-80 H Et

227-230 4.34(s, 3H), 4.77(s, 2H), 7.58(s, 1H) I-81 Me Et

161.5-162.5 3.72(s, 3H), 4.10(s, 2H) 4.90(s, 2H), 6.74 (s, 1H) I-82 H Et

200-201.5 4.23(s, 3H), 4.76(s, 2H), 7.48(s, 1H)

TABLE 10

Com- Melting ¹H-NMR: δ pound No. R³⁶ R³⁷ R³⁸ R³⁹ point (° C.) CDCl₃(R³⁶= Me), DMSO-d₆(R³⁶ = H) I-83 Me Me

Me 205-207 2.18(d, J=1.2Hz, 3H), 2.42(s, 3H), 3.74(s, 3H), 4.19(s, 2H),4.97 (s, 2H), 7.11(d, J=1.2 Hz, 1H) I-84 H Me

Me 199.5-201 2.19(d, J=0.9Hz, 3H), 2.34(s, 3H), 4.31(s, 2H), 4.81(s,2H), 7.77 (d, J=0.9Hz, 1H) I-85 Me Et

Me 203-204 2.23(d, J=1.2Hz, 3H), 2.40(s, 3H), 3.75(s, 3H), 4.25(s, 2H),4.97 (s, 2H), 7.22(d, J=1.2 Hz, 1H) I-86 H Et

Me 216.5-218 2.21(s, 3H), 2.32(s, 3H), 4.41(s, 2H), 4.80 (s, 2H),7.78(s, 1H) I-87 Me Et

Me 186-187 2.16(d, J=0.9Hz, 3H), 3.77(s, 3H), 4.32(d, J=1.5Hz, 2H),4.99(s, 2H), 6.61(s, 1H), 7.05 (d, J=0.9Hz, 1H) I-88 H Et

Me 211-213 2.17(s, 3H), 4.50(s, 2H), 4.82(s, 2H), 6.80 (s, 1H), 7.72(s,1H)

TABLE 11

Com- Melting pound No. R³⁶ R³⁸ point (° C.) ¹H-NMR(CDCI₃):δ I-89 Et

184-186 1.81(t, J=7.8Hz, 3H), 2.18(s, 3H), 4.17(s, 2H), 4.22(q, J=7.8Hz, 2H), 4.94(s, 2H), 6.96(s, 1H) I-90 Et

172-173 1.26(t, J=7.2Hz, 3H), 2.13(s, 3H), 4.05(s, 2H), 4.21(q, J=7.2Hz, 2H), 4.92(s, 2H), 6.75(s, 1H) I-91 Et

160-161 1.26(t, J=7.2Hz, 3H), 2.14(d, J=1.2Hz, 3H), 4.21(q, J=7.2 Hz,2H), 4.25(s, 2H), 4.93(s, 2H), 6.85(d, J=1.2Hz, 1H) I-92 Et

185-186 1.26(t, J=7.2Hz, 3H), 2.13(d, J=0.9Hz, 3H), 4.16(s, 2H), 4.22(q, J=7.2Hz, 2H), 4.92(s, 2H), 6.79(d, J=0.9Hz, 1H) I-93 Pr

182-183 0.89(t, J=7.5Hz, 3H), 1.63(m, 2H), 2.17(s, 3H), 4.12(t, J=6.6Hz, 2H), 4.17(s, 2H), 4.95(s, 2H), 6.97(s, 1H) I-94 Pentyl

175-176 0.88(t, J=6.9Hz, 3H), 1.29(m, 4H), 1.62(m, 2H), 2.17(s, 3H),4.15(t, J=6.6Hz, 2H), 4.17(s, 2H), 4.95(s, 2H), 6.97(s, 1H) I-95

139-140 2.19(s, 3H), 2.50(m, 4H), 2.66 (t, J=5.7Hz, 2H), 3.71(t, J=4.5Hz, 4H), 4.17(s, 2H), 4.30(t, J=5.7Hz, 2H), 5.03(s, 2H), 6.97 (s, 1H)I-96

2.14(s, 3H), 2.50(m, 4H), 2.66 (m, 2H), 3.70(m, 4H), 4.06(s, 2H),4.30(t, J=5.4Hz, 2H), 5.01 (s, 2H), 6.75(s, 1H)

TABLE 12 Com- Melting pound No. R³⁶ R³⁸ point (° C.) ¹H-NMR(CDCI₃):δI-97

2.15(s, 3H), 2.49(m, 4H), 2.65 (t, J=5.4Hz, 2H), 3.70(m. 4H), 4.26(s,2H), 4.29(t, J=5.4Hz, 2H), 5.01(s, 2H), 6.85(s, 1H) I-98

2.13(d, J=0.9Hz, 3H), 2.40- 2.80(m 6H), 3.66-3.78(m, 4H), 4.16(s, 2H),4.26-4.36(m, 2H), 5.01(s, 2H), 6.80(d, J=0.9Hz, 1H) I-99 Na

250-265 *2.18(s, 3H), 4.28(s, 2H), 4.81 (s, 2H), 7.31(s, 1H) I-100 Na

**2.14(d, J=0.9Hz, 3H), 4.14 (s, 2H), 4.79(s, 2H), 6.99(d, J=0.9Hz, 1H)I-101 Na

*2.15(d, J=0.9Hz, 3H), 4.32(s, 2H), 4.80(s, 2H), 7.05(d, J=0.9 Hz, 1H)I-102 Na

260-263 **2.12(d, J=0.9Hz, 3H), 4.25 (s, 2H), 4.42(s, 2H), 7.21(m, 1H)*measured with CD₃OD **measured with DMSO-d₆

TABLE 13

Com- Melting pound No. R³⁶ point (° C.) ¹H-NMR(CDCI₃):δ I-103

147-148 2.12(s, 3H), 2.18(s, 3H), 4.18 (s, 2H), 5.01(s, 2H), 5.81(s,2H), 6.97(s, 1H) I-104

143-144 1.20(s, 9H), 2.17(s, 3H), 4.17 (s, 2H), 5.01(s, 2H), 5.81(s,2H), 6.97(s, 1H) I-105

148-150 2.18(s, 3H), 3.83(s, 3H), 4.17 (s, 2H), 5.02(s, 2H), 5.83(s,2H), 6.97(s, 1H) I-106

125-130 1.10-2.00(m, 10H), 1.53(d, J=5.7Hz, 3H), 2.18(s, 3H), 4.17 (s,2H), 4.65(m, 1H), 4.94(d, J=15.9Hz, 1H), 5.00(d, J=15.9 Hz, 1H), 6.82(q,J=5.7Hz, 1H), 6.98(s, 1H)

The compounds shown in the following Tables 14 to 25 can be synthesizedin accordance with the same method describe in the above Examples. Theabbreviations used in Tables 14 to 25: AA, AB, AC, AD, AE, AF, AG, BA,BB, BC, BD, BE, BF, BG, BH, BI, BJ, BK, BL, BM, BN, BO, BP, BQ, BR, BS,BT, BU, BV, BW, BX, BY, BZ, CA, CB, CC, CD, CE, CF, CG, CH, CI, CJ, CK,CL, CM, and CN show the substituents described as follows.

AA

AB

AC

AD

AE

AF

AG

BA

BB

BC

BD

BE

BF

BG

BH

BI

BJ

BK

BL

BM

BN

BO

BP

BQ

BR

BS

BT

BU

BV

BW

BX

BY

BZ

CA

CB

CC

CD

CE

CF

CG

CH

CI

CJ

CK

CL

CM

CN

TABLE 14

Com- pound No. R³⁷ R³⁸ R³⁹ II-1 Me BA Me II-2 Me BB Me II-3 Me BC MeII-4 Me BD Me II-5 Me BE Me II-6 Me BF Me II-7 Me BG Me II-8 Me BH MeII-9 Me BI Me II-10 Me BJ Me II-11 Me BK Me II-12 Me BL Me II-13 Me BMMe II-14 Me BN Me II-15 Me BO Me II-16 Me BP Me II-17 Me BQ Me II-18 MeBR Me II-19 Me BS Me II-20 Me BT Me II-21 Me BU Me II-22 Me BV Me II-23Me BW Me II-24 Me BX Me II-25 Me BY Me II-26 Me BZ Me II-27 Me CA MeII-28 Me CB Me II-29 Me CC Me II-30 Me CD Me II-31 Me CE Me II-32 Me CFMe II-33 Me CG Me II-34 Me CH Me II-35 Me CI Me II-36 Me CJ Me II-37 MeCK Me II-38 Me CL Me II-39 Me CM Me II-40 Me CN Me II-41 Et BA Me II-42Et BB Me II-43 Et BC Me II-44 Et BD Me II-45 Et BE Me II-46 Et BF MeII-47 Et BG Me II-48 Et BH Me II-49 Et BI Me II-50 Et BJ Me II-51 Et BKMe II-52 Et BL Me II-53 Et BM Me II-54 Et BN Me II-55 Et BO Me II-56 EtBP Me II-57 Et BQ Me II-58 Et BR Me II-59 Et BS Me II-60 Et BT Me II-61Et BU Me II-62 Et BV Me II-63 Et BW Me II-64 Et BX Me II-65 Et BY MeII-66 Et BZ Me II-67 Et CA Me II-68 Et CB Me II-69 Et CC Me II-70 Et CDMe II-71 Et CE Me II-72 Et CF Me II-73 Et CG Me II-74 Et CH Me II-75 EtCI Me II-76 Et CJ Me II-77 Et CK Me II-78 Et CL Me II-79 Et CM Me II-80Et CN Me II-81 Ph BA Me II-82 Ph BB Me II-83 Ph BC Me II-84 Ph BD MeII-85 Ph BE Me II-86 Ph BF Me II-87 Ph BG Me II-88 Ph BH Me II-89 Ph BIMe II-90 Ph BJ Me II-91 Ph BK Me II-92 Ph BL Me II-93 Ph BM Me II-94 PhBN Me II-95 Ph BO Me II-96 Ph BP Me II-97 Ph BQ Me II-98 Ph BR Me II-99Ph BS Me II-100 Ph BT Me II-101 Ph BU Me II-102 Ph BV Me II-103 Ph BW MeII-104 Ph BX Me II-105 Ph BY Me II-106 Ph BZ Me II-107 Ph CA Me II-108Ph CB Me II-109 Ph CC Me II-110 Ph CD Me II-111 Ph CE Me II-112 Ph CF MeII-113 Ph CG Me II-114 Ph CH Me II-115 Ph CI Me II-116 Ph CJ Me II-117Ph CK Me II-118 Ph CL Me II-119 Ph CM Me II-120 Ph CN Me

TABLE 15

Com- pound No. R³⁷ R³⁸ R³⁹ II-121 Me BA Et II-122 Me BB Et II-123 Me BCEt II-124 Me BD Et II-125 Me BE Et II-126 Me BF Et II-127 Me BG EtII-128 Me BH Et II-129 Me BI Et II-130 Me BJ Et II-131 Me BK Et II-132Me BL Et II-133 Me BM Et II-134 Me BN Et II-135 Me BO Et II-136 Me BP EtII-137 Me BQ Et II-138 Me BR Et II-139 Me BS Et II-140 Me BT Et II-141Me BU Et II-142 Me BV Et II-143 Me BW Et II-144 Me BX Et II-145 Me BY EtII-146 Me BZ Et II-147 Me CA Et II-148 Me CB Et II-149 Me CC Et II-150Me CD Et II-151 Me CE Et II-152 Me CF Et II-153 Me CG Et II-154 Me CH EtII-155 Me CI Et II-156 Me CJ Et II-157 Me CK Et II-158 Me CL Et II-159Me CM Et II-160 Me CN Et II-161 Et BA Et II-162 Et BB Et II-163 Et BC EtII-164 Et BD Et II-165 Et BE Et II-166 Et BF Et II-167 Et BG Et II-168Et BH Et II-169 Et BI Et II-170 Et BJ Et II-171 Et BK Et II-172 Et BL EtII-173 Et BM Et II-174 Et BN Et II-175 Et BO Et II-176 Et BP Et II-177Et BQ Et II-178 Et BR Et II-179 Et BS Et II-180 Et BT Et II-181 Et BU EtII-182 Et BV Et II-183 Et BW Et II-184 Et BX Et II-185 Et BY Et II-186Et BZ Et II-187 Et CA Et II-188 Et CB Et II-189 Et CC Et II-190 Et CD EtII-191 Et CE Et II-192 Et CF Et II-193 Et CG Et II-194 Et CH Et II-195Et CI Et II-196 Et CJ Et II-197 Et CK Et II-198 Et CL Et II-199 Et CM EtII-200 Et CN Et II-201 Ph BA Et II-202 Ph BB Et II-203 Ph BC Et II-204Ph BD Et II-205 Ph BE Et II-206 Ph BF Et II-207 Ph BG Et II-208 Ph BH EtII-209 Ph BI Et II-210 Ph BJ Et II-211 Ph BK Et II-212 Ph BL Et II-213Ph BM Et II-214 Ph BN Et II-215 Ph BO Et II-216 Ph BP Et II-217 Ph BQ EtII-218 Ph BR Et II-219 Ph BS Et II-220 Ph BT Et II-221 Ph BU Et II-222Ph BV Et II-223 Ph BW Et II-224 Ph BX Et II-225 Ph BY Et II-226 Ph BZ EtII-227 Ph CA Et II-228 Ph CB Et II-229 Ph CC Et II-230 Ph CD Et II-231Ph CE Et II-232 Ph CF Et II-233 Ph CG Et II-234 Ph CH Et II-235 Ph CI EtII-236 Ph CJ Et II-237 Ph CK Et II-238 Ph CL Et II-239 Ph CM Et II-240Ph CN Et

TABLE 16

Com- pound No. R³⁷ R³⁸ R³⁹ II-241 Me BA Me II-242 Me BB Me II-243 Me BCMe II-244 Me BD Me II-245 Me BE Me II-246 Me BF Me II-247 Me BG MeII-248 Me BH Me II-249 Me BI Me II-250 Me BJ Me II-251 Me BK Me II-252Me BL Me II-253 Me BM Me II-254 Me BN Me II-255 Me BO Me II-256 Me BP MeII-257 Me BQ Me II-258 Me BR Me II-259 Me BS Me II-260 Me BT Me II-261Me BU Me II-262 Me BV Me II-263 Me BW Me II-264 Me BX Me II-265 Me BY MeII-266 Me BZ Me II-267 Me CA Me II-268 Me CB Me II-269 Me CC Me II-270Me CD Me II-271 Me CE Me II-272 Me CF Me II-273 Me CG Me II-274 Me CH MeII-275 Me CI Me II-276 Me CJ Me II-277 Me CK Me II-278 Me CL Me II-279Me CM Me II-280 Me CN Me II-281 Et BA Me II-282 Et BB Me II-283 Et BC MeII-284 Et BD Me II-285 Et BE Me II-286 Et BF Me II-287 Et BG Me II-288Et BH Me II-289 Et BI Me II-290 Et BJ Me II-291 Et BK Me II-292 Et BL MeII-293 Et BM Me II-294 Et BN Me II-295 Et BO Me II-296 Et BP Me II-297Et BQ Me II-298 Et BR Me II-299 Et BS Me II-300 Et BT Me II-301 Et BU MeII-302 Et BV Me II-303 Et BW Me II-304 Et BX Me II-305 Et BY Me II-306Et BZ Me II-307 Et CA Me II-308 Et CB Me II-309 Et CC Me II-310 Et CD MeII-311 Et CE Me II-312 Et CF Me II-313 Et CG Me II-314 Et CH Me II-315Et CI Me II-316 Et CJ Me II-317 Et CK Me II-318 Et CL Me II-319 Et CM MeII-320 Et CN Me II-321 Ph BA Me II-322 Ph BB Me II-323 Ph BC Me II-324Ph BD Me II-325 Ph BE Me II-326 Ph BF Me II-327 Ph BG Me II-328 Ph BH MeII-329 Ph BI Me II-330 Ph BJ Me II-331 Ph BK Me II-332 Ph BL Me II-333Ph BM Me II-334 Ph BN Me II-335 Ph BO Me II-336 Ph BP Me II-337 Ph BQ MeII-338 Ph BR Me II-339 Ph BS Me II-340 Ph BT Me II-341 Ph BU Me II-342Ph BV Me II-343 Ph BW Me II-344 Ph BX Me II-345 Ph BY Me II-346 Ph BZ MeII-347 Ph CA Me II-348 Ph CB Me II-349 Ph CC Me II-350 Ph CD Me II-351Ph CE Me II-352 Ph CF Me II-353 Ph CG Me II-354 Ph CH Me II-355 Ph CI MeII-356 Ph CJ Me II-357 Ph CK Me II-358 Ph CL Me II-359 Ph CM Me II-360Ph CN Me

TABLE 17

Com- pound No. R³⁷ R³⁸ R³⁹ II-361 Me BA Et II-362 Me BB Et II-363 Me BCEt II-364 Me BD Et II-365 Me BE Et II-366 Me BF Et II-367 Me BG EtII-368 Me BH Et II-369 Me BI Et II-370 Me BJ Et II-371 Me BK Et II-372Me BL Et II-373 Me BM Et II-374 Me BN Et II-375 Me BO Et II-376 Me BP EtII-377 Me BQ Et II-378 Me BR Et II-379 Me BS Et II-380 Me BT Et II-381Me BU Et II-382 Me BV Et II-383 Me BW Et II-384 Me BX Et II-385 Me BY EtII-386 Me BZ Et II-387 Me CA Et II-388 Me CB Et II-389 Me CC Et II-390Me CD Et II-391 Me CE Et II-392 Me CF Et II-393 Me CG Et II-394 Me CH EtII-395 Me CI Et II-396 Me CJ Et II-397 Me CK Et II-398 Me CL Et II-399Me CM Et II-400 Me CN Et II-401 Et BA Et II-402 Et BB Et II-403 Et BC EtII-404 Et BD Et II-405 Et BE Et II-406 Et BF Et II-407 Et BG Et II-408Et BH Et II-409 Et BI Et II-410 Et BJ Et II-411 Et BK Et II-412 Et BL EtII-413 Et BM Et II-414 Et BN Et II-415 Et BO Et II-416 Et BP Et II-417Et BQ Et II-418 Et BR Et II-419 Et BS Et II-420 Et BT Et II-421 Et BU EtII-422 Et BV Et II-423 Et BW Et II-424 Et BX Et II-425 Et BY Et II-426Et BZ Et II-427 Et CA Et II-428 Et CB Et II-429 Et CC Et II-430 Et CD EtII-431 Et CE Et II-432 Et CF Et II-433 Et CG Et II-434 Et CH Et II-435Et CI Et II-436 Et CJ Et II-437 Et CK Et II-438 Et CL Et II-439 Et CM EtII-440 Et CN Et II-441 Ph BA Et II-442 Ph BB Et II-443 Ph BC Et II-444Ph BD Et II-445 Ph BE Et II-446 Ph BF Et II-447 Ph BG Et II-448 Ph BH EtII-449 Ph BI Et II-450 Ph BJ Et II-451 Ph BK Et II-452 Ph BL Et II-453Ph BM Et II-454 Ph BN Et II-455 Ph BO Et II-456 Ph BP Et II-457 Ph BQ EtII-458 Ph BR Et II-459 Ph BS Et II-460 Ph BT Et II-461 Ph BU Et II-462Ph BV Et II-463 Ph BW Et II-464 Ph BX Et II-465 Ph BY Et II-466 Ph BZ EtII-467 Ph CA Et II-468 Ph CB Et II-469 Ph CC Et II-470 Ph CD Et II-471Ph CE Et II-472 Ph CF Et II-473 Ph CG Et II-474 Ph CH Et II-475 Ph CI EtII-476 Ph CJ Et II-477 Ph CK Et II-478 Ph CL Et II-479 Ph CM Et II-480Ph CN Et

TABLE 18

Com- pound No. R³⁷ R³⁸ R³⁹ II-481 Me BA Me II-482 Me BB Me II-483 Me BCMe II-484 Me BD Me II-485 Me BE Me II-486 Me BF Me II-487 Me BG MeII-488 Me BH Me II-489 Me BI Me II-490 Me BJ Me II-491 Me BK Me II-492Me BL Me II-493 Me BM Me II-494 Me BN Me II-495 Me BO Me II-496 Me BP MeII-497 Me BQ Me II-498 Me BR Me II-499 Me BS Me II-490 Me BT Me II-501Me BU Me II-502 Me BV Me II-503 Ne BW Me II-504 Me BX Me II-505 Me BY MeII-506 Me BZ Me II-507 Me CA Me II-508 Me CB Me II-509 Me CC Me II-510Me CD Me II-511 Me CE Me II-512 Me CF Me II-513 Me CG Me II-514 Me CH MeII-515 Me CI Me II-516 Me CJ Me II-517 Me CK Me II-518 Me CL Me II-519Me CM Me II-520 Me CN Me II-521 Et BA Me II-522 Et BB Me II-523 Et BC MeII-524 Et BD Me II-525 Et BE Me II-526 Et BF Me II-527 Et BG Me II-528Et BH Me II-529 Et BI Me II-530 Et BJ Me II-531 Et BK Me II-532 Et BL MeII-533 Et BM Me II-534 Et BN Me II-535 Et BO Me II-536 Et BP Me II-537Et BQ Me II-538 Et BR Me II-539 Et BS Me II-540 Et BT Me II-54J Et BU MeII-542 Et BV Me II-543 Et BW Me II-544 Et BX Me II-545 Et BY Me II-546Et BZ Me II-547 Et CA Me II-548 Et CB Me II-549 Et CC Me II-550 Et CD MeII-551 Et CE Me II-552 Et CF Me II-553 Et CG Me II-554 Et CH Me II-555Et CI Me II-556 Et CJ Me II-557 Et CK Me II-558 Et CL Me II-559 Et CM MeII-560 Et CN Me II-561 Ph BA Me II-562 Ph BB Me II-563 Ph BC Me II-564Ph BD Me II-565 Ph BE Me II-566 Ph BF Me II-567 Ph BG Me II-568 Ph BH MeII-569 Ph BI Me II-570 Ph BJ Me II-57J Ph BK Me II-572 Ph BL Me II-573Ph BM Me II-574 Ph BN Me II-575 Ph BO Me II-576 Ph BP Me II-577 Ph BQ MeII-578 Ph BR Me II-579 Ph BS Me II-580 Ph BT Me II-581 Ph BU Me II-582Ph BV Me II-583 Ph BW Me II-584 Ph BX Me II-585 Ph BY Me II-586 Ph BZ MeII-587 Ph CA Me II-588 Ph CB Me II-589 Ph CC Me II-590 Ph CD Me II-591Ph CE Me II-592 Ph CF Me II-593 Ph CG Me II-594 Ph CH Me II-595 Ph CI MeII-596 Ph CJ Me II-597 Ph CK Me II-598 Ph CL Me II-599 Ph CM Me II-600Ph CN Me

TABLE 19

Com- pound No. R³⁷ R³⁸ R³⁹ II-601 Me BA Et II-602 Me BB Et II-603 Me BCEt II-604 Me BD Et II-605 Me BE Et II-606 Me BF Et II-607 Me BG EtII-608 Me BH Et II-609 Me BI Et II-610 Me BJ Et II-611 Me BK Et II-612Me BL Et II-613 Me BM Et II-614 Me BN Et II-615 Me BO Et II-616 Me BP EtII-617 Me BQ Et II-618 Me BR Et II-619 Me BS Et II-620 Me BT Et II-621Me BU Et II-622 Me BV Et II-623 Me BW Et II-624 Me BX Et II-625 Me BY EtII-626 Me BZ Et II-627 Me CA Et II-628 Me CB Et II-629 Me CC Et II-630Me CD Et II-631 Me CE Et II-632 Me CF Et II-633 Me CG Et II-634 Me CH EtII-635 Me CI Et II-636 Me CJ Et II-637 Me CK Et II-638 Me CL Et II-639Me CM Et II-640 Me CN Et II-641 Et BA Et II-642 Et BB Et II-643 Et BC EtII-644 Et BD Et II-645 Et BE Et II-646 Et BF Et II-647 Et BG Et II-648Et BH Et II-649 Et BI Et II-650 Et BJ Et II-651 Et BK Et II-652 Et BL EtII-653 Et BM Et II-654 Et BN Et II-655 Et BO Et II-656 Et BP Et II-657Et BQ Et II-658 Et BR Et II-659 Et BS Et II-660 Et BT Et II-661 Et BU EtII-662 Et BV Et II-663 Et BW Et II-664 Et BX Et II-665 Et BY Et II-666Et BZ Et II-667 Et CA Et II-668 Et CB Et II-669 Et CC Et II-670 Et CD EtII-671 Et CE Et II-672 Et CF Et II-673 Et CG Et II-674 Et CH Et II-675Et CI Et II-676 Et CJ Et II-677 Et CK Et II-678 Et CL Et II-679 Et CM EtII-680 Et CN Et II-681 Ph BA Et II-682 Ph BB Et II-683 Ph BC Et II-684Ph BD Et II-685 Ph BE Et II-686 Ph BF Et II-687 Ph BG Et II-688 Ph BH EtII-689 Ph BI Et II-690 Ph BJ Et II-691 Ph BK Et II-692 Ph BL Et II-693Ph BM Et II-694 Ph BN Et II-695 Ph BO Et II-696 Ph BP Et II-697 Ph BQ EtII-698 Ph BR Et II-699 Ph BS Et II-700 Ph BT Et II-701 Ph BU Et II-702Ph BV Et II-703 Ph BW Et II-704 Ph BX Et II-705 Ph BY Et II-706 Ph BZ EtII-707 Ph CA Et II-708 Ph CB Et II-709 Ph CC Et II-710 Ph CD Et II-711Ph CE Et II-712 Ph CF Et II-713 Ph CG Et II-714 Ph CH Et II-715 Ph CI EtII-716 Ph CJ Et II-717 Ph CK Et II-718 Ph CL Et II-719 Ph CM Et II-720Ph CN Et

TABLE 20

Compound No. R³⁷ R³⁹ R⁴⁰ R⁴¹ Compound No. R³⁷ R³⁹ R⁴⁰ R⁴¹ III-1  Me MeAA H III-36 Me Et AA Et III-2  Me Me AB H III-37 Me Et AB Et III-3  MeMe AC H III-38 Me Et AC Et III-4  Me Me AD H III-39 Me Et AD Et III-5 Me Me AE H III-40 Me Et AE Et III-6  Me Me AF H III-41 Me Et AF EtIII-7  Me Me AG H III-42 Me Et AG Et III-8  Me Me AA Me III-43 Me Ph AAH III-9  Me Me AB Me III-44 Me Ph AB H III-10 Me Me AC Me III-45 Me PhAC H III-11 Me Me AD Me III-46 Me Ph AD H III-12 Me Me AE Me III-47 MePh AE H III-13 Me Me AF Me III-48 Me Ph AF H III-14 Me Me AG Me III-49Me Ph AG H III-15 Me Me AA Et III-50 Me Ph AA Me III-16 Me Me AB EtIII-51 Me Ph AB Me III-17 Me Me AC Et III-52 Me Pb AC Me III-18 Me Me ADEt III-53 Me Ph AD Me III-19 Me Me AE Et III-54 Me Ph AE Me III-20 Me MeAF Et III-55 Me Ph AF Me III-21 Me Me AG Et III-56 Me Ph AG Me III-22 MeEt AA H III-57 Me Ph AA Et III-23 Me Et AB H III-58 Me Ph AB Et III-24Me Et AC H III-59 Me Ph AC Et III-25 Me Et AD H III-60 Me Ph AD EtIII-26 Me Et AE H III-61 Me Ph AE Et III-27 Me Et AF H III-62 Me Ph AFEt III-28 Me Et AG H III-63 Me Ph AG Et III-29 Me Et AA Me III-30 Me EtAB Me III-31 Me Et AC Me III-32 Me Et AD Me III-33 Me Et AE Me III-34 MeEt AF Me III-35 Me Et AG Me

TABLE 21

Compound No. R³⁷ R³⁹ R⁴⁰ R⁴¹ Compound No. R³⁷ R³⁹ R⁴⁰ R⁴¹ III-64 Et MeAA H III-99  Et Et AA Et III-65 Et Me AB H III-100 Et Et AB Et III-66 EtMe AC H III-101 Et Et AC Et III-67 Et Me AD H III-102 Et Et AD Et III-68Et Me AE H III-103 Et Et AE Et III-69 Et Me AF H III-104 Et Et AF EtIII-70 Et Me AG H III-105 Et Et AG Et III-71 Et Me AA Me III-106 Et PhAA H III-72 Et Me AB Me III-107 Et Ph AB H III-73 Et Me AC Me III-108 EtPh AC H III-74 Et Me AD Me III-109 Et Ph AD H III-75 Et Me AE Me III-110Et Ph AE H III-76 Et Me AF Me III-111 Et Ph AF H III-77 Et Me AG MeIII-112 Et Ph AG H III-78 Et Me AA Et III-113 Et Ph AA Me III-79 Et MeAB Et III-114 Et Ph AB Me III-80 Et Me AC Et III-115 Et Ph AC Me III-81Et Me AD Et III-116 Et Ph AD Me III-82 Et Me AE Et III-117 Et Ph AE MeIII-83 Et Me AF Et III-118 Et Ph AF Me III-84 Et Me AG Et III-119 Et PhAG Me III-85 Et Et AA H III-120 Et Ph AA Et III-86 Et Et AB H III-121 EtPh AB Et III-87 Et Et AC H III-122 Et Ph AC Et III-88 Et Et AD H III-123Et Ph AD Et III-89 Et Et AE H III-124 Et Ph AE Et III-90 Et Et AF HIII-125 Et Ph AF Et III-91 Et Et AG H III-126 Et Ph AG Et III-92 Et EtAA Me III-93 Et Et AB Me III-94 Et Et AC Me III-95 Et Et AD Me III-96 EtEt AE Me III-97 Et Et AF Me III-98 Et Et AG Me

TABLE 22

Compound No. R³⁷ R³⁹ R⁴⁰ R⁴¹ Compound No. R³⁷ R³⁹ R⁴⁰ R⁴¹ III-127 Me MeAA H III-162 Me Et AA Et III-128 Me Me AB H III-163 Me Et AB Et III-129Me Me AC H III-164 Me Et AC Et III-130 Me Me AD H III-165 Me Et AD EtIII-131 Me Me AE H III-166 Me Et AE Et III-132 Me Me AF H III-167 Me EtAF Et III-133 Me Me AG H III-168 Me Et AG Et III-134 Me Me AA Me III-169Me Ph AA H III-135 Me Me AB Me III-170 Me Ph AB H III-136 Me Me AC MeIII-171 Me Ph AC H III-137 Me Me AD Me III-172 Me Ph AD H III-138 Me MeAE Me III-173 Me Ph AE H III-139 Me Me AF Me III-174 Me Ph AF H III-140Me Me AG Me III-175 Me Ph AG H III-141 Me Me AA Et III-176 Me Ph AA MeIII-142 Me Me AB Et III-177 Me Ph AB Me III-143 Me Me AC Et III-178 MePh AC Me III-144 Me Me AD Et III-179 Me Ph AD Me III-145 Me Me AE EtIII-180 Me Ph AE Me III-146 Me Me AF Et III-181 Me Ph AF Me III-147 MeMe AG Et III-182 Me Ph AG Me III-148 Me Et AA H III-183 Me Ph AA EtIII-149 Me Et AB H III-184 Me Ph AB Et III-150 Me Et AC H III-185 Me PhAC Et III-151 Me Et AD H III-186 Me Ph AD Et III-152 Me Et AE H III-187Me Ph AE Et III-153 Me Et AF H III-188 Me Ph AF Et III-154 Me Et AG HIII-189 Me Ph AG Et III-155 Me Et AA Me III-156 Me Et AB Me III-157 MeEt AC Me III-158 Me Et AD Me III-159 Me Et AE Me III-160 Me Et AF MeIII-161 Me Et AG Me

TABLE 23

Compound No. R³⁷ R³⁹ R⁴⁰ R⁴¹ Compound No. R³⁷ R³⁹ R⁴⁰ R⁴¹ III-190 Et MeAA H III-225 Et Et AA Et III-191 Et Me AB H III-226 Et Et AB Et III-192Et Me AC H III-227 Et Et AC Et III-193 Et Me AD H III-228 Et Et AD EtIII-194 Et Me AE H III-229 Et Et AE Et III-195 Et Me AF H III-230 Et EtAF Et III-196 Et Me AG H III-231 Et Et AG Et III-197 Et Me AA Me III-232Et Ph AA H III-198 Et Me AB Me III-233 Et Ph AB H III-199 Et Me AC MeIII-234 Et Ph AC H III-200 Et Me AD Me III-235 Et Ph AD H III-201 Et MeAE Me III-236 Et Ph AE H III-202 Et Me AF Me III-237 Et Ph AF H III-203Et Me AG Me III-238 Et Ph AG H III-204 Et Me AA Et III-239 Et Ph AA MeIII-205 Et Me AB Et III-240 Et Ph AB Me III-206 Et Me AC Et III-241 EtPh AC Me III-207 Et Me AD Et III-242 Et Ph AD Me III-208 Et Me AE EtIII-243 Et Ph AE Me III-209 Et Me AF Et III-244 Et Ph AF Me III-210 EtMe AG Et III-245 Et Ph AG Me III-211 Et Et AA H III-246 Et Ph AA EtIII-212 Et Et AB H III-247 Et Ph AB Et III-213 Et Et AC H III-248 Et PhAC Et III-214 Et Et AD H III-249 Et Ph AD Et III-215 Et Et AE H III-250Et Ph AE Et III-216 Et Et AF H III-251 Et Ph AF Et III-217 Et Et AG HIII-252 Et Ph AG Et III-218 Et Et AA Me III-219 Et Et AB Me III-220 EtEt AC Me III-221 Et Et AD Me III-222 Et Et AE Me III-223 Et Et AF MeIII-224 Et Et AG Me

TABLE 24

Compound No. R³⁷ R³⁹ R⁴⁰ R⁴¹ Compound No. R³⁷ R³⁹ R⁴⁰ R⁴¹ III-253 Me MeAA H III-288 Me Et AA Et III-254 Me Me AB H III-289 Me Et AB Et III-255Me Me AC H III-290 Me Et AC Et III-256 Me Me AD H III-291 Me Et AD EtIII-257 Me Me AE H III-292 Me Et AE Et III-258 Me Me AF H III-293 Me EtAF Et III-259 Me Me AG H III-294 Me Et AG Et III-260 Me Me AA Me III-295Me Ph AA H III-261 Me Me AB Me III-296 Me Ph AB H III-262 Me Me AC MeIII-297 Me Ph AC H III-263 Me Me AD Me III-298 Me Ph AD H III-264 Me MeAE Me III-299 Me Ph AE H III-265 Me Me AF Me III-300 Me Ph AF H III-266Me Me AG Me III-301 Me Ph AG H III-267 Me Me AA Et III-302 Me Ph AA MeIII-268 Me Me AB Et III-303 Me Ph AB Me III-269 Me Me AC Et III-304 MePh AC Me III-270 Me Me AD Et III-305 Me Ph AD Me III-271 Me Me AE EtIII-306 Me Ph AE Me III-272 Me Me AF Et III-307 Me Ph AF Me III-273 MeMe AG Et III-308 Me Ph AG Me III-274 Me Et AA H III-309 Me Ph AA EtIII-275 Me Et AB H III-310 Me Ph AB Et III-276 Me Et AC H III-311 Me PhAC Et III-277 Me Et AD H III-312 Me Ph AD Et III-278 Me Et AE H III-313Me Ph AE Et III-279 Me Et AF H III-314 Me Ph AF Et III-280 Me Et AG HIII-315 Me Ph AG Et III-281 Me Et AA Me III-282 Me Et AB Me III-283 MeEt AC Me III-284 Me Et AD Me III-285 Me Et AE Me III-286 Me Et AF MeIII-287 Me Et AG Me

TABLE 25

Compound No. R³⁷ R³⁹ R⁴⁰ R⁴¹ Compound No. R³⁷ R³⁹ R⁴⁰ R⁴¹ III-316 Et MeAA H III-351 Et Et AA Et III-317 Et Me AB H III-352 Et Et AB Et III-318Et Me AC H III-353 Et Et AC Et III-319 Et Me AD H III-354 Et Et AD EtIII-320 Et Me AE H III-355 Et Et AE Et III-321 Et Me AF H III-356 Et EtAF Et III-322 Et Me AG H III-357 Et Et AG Et III-323 Et Me AA Me III-358Et Ph AA H III-324 Et Me AB Me III-359 Et Ph AB H III-325 Et Me AC MeIII-360 Et Ph AC H III-326 Et Me AD Me III-361 Et Ph AD H III-327 Et MeAE Me III-362 Et Ph AE H III-328 Et Me AF Me III-363 Et Ph AF H III-329Et Me AG Me III-364 Et Ph AG H III-330 Et Me AA Et III-365 Et Ph AA MeIII-331 Et Me AB Et III-366 Et Ph AB Me III-332 Et Me AC Et III-367 EtPh AC Me III-333 Et Me AD Et III-368 Et Ph AD Me III-334 Et Me AE EtIII-369 Et Ph AE Me III-335 Et Me AF Et III-370 Et Ph AF Me III-336 EtMe AG Et III-371 Et Ph AG Me III-337 Et Et AA H III-372 Et Ph AA EtIII-338 Et Et AB H III-373 Et Ph AB Et III-339 Et Et AC H III-374 Et PhAC Et III-340 Et Et AD H III-375 Et Ph AD Et III-341 Et Et AE H III-376Et Ph AE Et III-342 Et Et AF H III-377 Et Ph AF Et III-343 Et Et AG HIII-378 Et Ph AG Et III-344 Et Et AA Me III-345 Et Et AB Me III-346 EtEt AC Me III-347 Et Et AD Me III-348 Et Et AE Me III-349 Et Et AF MeIII-350 Et Et AG Me

Test Example: Inhibition Test of Human Secretory Phospholipase A₂

Analytical Experiment

In order to identify and evaluate an inhibitor of recombinant humansecretory phospholipase A₂, the following chromogenic assay is utilized.The assay herein has been applied for high volume screening wherein 96well microtiterplate is used. A general explanation for such assay isdescribed in “Analysis of Human Synovial Fluid Phospholipase A₂ on ShortChain Phosphatidylcholine-Mixed Micelles: Development of aSpectrophotometric Assay Suitable for a Micortiterplate Reader”(Analytical Biochemistry, 204, pp 190-197, 1992 by Laure. J. Reynolds.Lori L. Hughes and Edward A. Dennis: the disclosure of which isincorporated herein for reference.

Reagents:

Reaction Buffer-

CaCl₂.6H₂O (2.19 g/L) KCl (7.455 g/L) Bovine Serum Albumin (fatty acidfree) (1 g/L) (Sigma A-7030) Tris-HCl (3.94 g/L)

pH 7.5 (adjusted with NaOH)

Enzyme Buffer-

0.05 M-AcONa

0.2 M-NaCl

pH 4.5 (adjusted with acetic acid)

Enzyme Solution-

1 mg of sPLA₂ is dissolved in 1 ml of an enzyme buffer. Thereafter, thesolution is maintained at 4° C.

In the assay, 5 μl of the solution is diluted with 1995 μl of thereaction buffer to be used.

DTNB-

198 mg of 5,5′-dithiobis-2-benzoic acid (manufactured by Wako PureChemicals) is dissolved in 100 ml of H₂O

H 7.5 (adjusted with NaOH)

Substrate Solution-

100 mg of racemic1,2-bis(heptanoylthio)-1,2-dideoxy-sn-glycero-3-phospholylcholine isdissolved in 1 ml of chloroform.

Triton-X 100-

624.9 mg of Triton-X 100 is dissolved in the reaction buffer.

Enzyme Reaction: for 1 Plate of Microtiterplate

1) 0.106 ml of the substrate solution is put in a centrifugal tube, andnitrogen gas is jetted to remove the solvent. 0.54 ml of Triton-X 100 isadded thereto, the mixture is stirred, thereafter it is sonified in abath type sonification to dissolve. To the resulting product are added17.8 ml of the reaction buffer and 0.46 ml of DTNB, and 0.18 ml each ofthe admixture is poured to wells of the 96 well microtiterplate.

2) 10 μl of a test compound (or solvent blank) are added in accordancewith alignment of plates which has been previously set.

3) Incubation is effected at 40° C. for 15 minutes.

4) 20μl of an enzyme solution (sPLA₂) which has been previously diluted(50 ng/well) are added to start reaction (40° C., 30 minutes).

5) Changes in absorbancy for 30 minutes are measured by a plate reader,and inhibition activity was calculated (OD: 405 nm).

6) IC₅₀ was determined by plotting log concentration with respect toinhibition values within 10% to 90% inhibiting range.

Results of the human secretory phospholipase A, inhibition test areshown in the following Table 26.

TABLE 26 Compound NO. IC₅₀ (μM) I-1 0.208 I-2 0.011 I-3 2.623 I-4 0.035I-5 0.314 I-6 0.009 I-7 0.389 I-8 0.011 I-9 0.435 I-10 0.014 I-11 0.194I-12 0.010 I-13 0.157 I-14 0.011 I-15 0.512 I-16 0.006 I-17 0.172 I-180.009 I-19 0.562 I-20 0.021 I-21 0.041 I-22 0.008 I-23 0.651 I-24 0.017I-25 0.196 I-26 0.012 I-27 0.022 I-28 0.007 I-29 0.056 I-30 0.008 I-311.168 I-32 0.028 I-33 0.703 I-34 0.026 I-35 0.182 I-36 0.011 I-37 0.726I-38 0.033 I-39 0.151 I-40 0.012 I-41 0.107 I-42 0.010 I-43 0.041 I-440.007 I-45 0.117 I-46 0.010 I-47 0.389 I-48 0.015 I-49 0.211 I-50 0.017I-51 0.061 I-52 0.005 I-53 0.059 I-54 0.006 I-55 0.032 I-56 0.006 I-580.025 I-59 15.8 I-60 1.21 I-61 0.081 I-62 0.006 I-63 0.057 I-64 0.006I-65 1.55 I-66 0.045 I-67 0.057 I-68 0.008 I-69 0.033 I-70 0.005 I-710.901 I-72 0.013 I-73 0.129 I-74 0.006 I-75 1.46 I-76 0.029 I-77 1.38I-78 0.060 I-79 0.062 I-80 0.006 I-81 0.201 I-82 0.005 I-83 0.116 I-840.008 I-85 0.370 I-86 0.011 I-87 0.129 I-88 0.008 I-89 0.315 I-90 0.038I-91 0.048 I-92 0.076 I-93 0.282 I-94 0.650 I-95 0.175 I-96 0.077 I-970.078 I-98 0.102 I-99 0.021 I-100 0.021 I-101 0.019 I-102 0.020 I-1030.540 I-104 0.988 I-105 0.400 I-106 0.819

Formulation Example

It is to be noted that the following Formulation Examples 1 to 8 aremere illustration, but not intended to limit the scope of the invention.The term “active ingredient” means the compounds represented by theformula (I), the prodrugs thereof, their pharmaceutical acceptablesalts, or their solvates.

Formulation Example 1

Hard gelatin capsules are prepared using of the following ingredients:

Dose (mg/capsule) Active ingredient 250 Starch, dried 200 Magnesiumstearate  10 Total 460 mg

Formulation Example 2

A tablet is prepared using of the following ingredients:

Dose (mg/tablet) Active ingredient 250 Cellulose, microcrystalline 400Silicon dioxide, fumed  10 Stearic acid  5 Total 665 mg

The components are blended and compressed to form tablets each weighing665 mg.

Formulation Example 3

An aerosol solution is prepared containing the following components:

Weight Active ingredient 0.25 Ethanol 25.75 Propellant 22(chlorodifluoromethane) 74.00 Total 100.00

The active compound is mixed with ethanol and the admixture added to aportion of the propellant 22, cooled to −30° C. and transferred tofilling device. The required amount is then fed to stainless steelcontainer and diluted with the reminder of the propellant. The valveunits are then fitted to the container.

Formulation Example 4

Tablets, each containing 60 mg of active ingredient, are made asfollows.

Active ingredient 60 mg Starch 45 mg Microcrystalline cellulose 35 mgPolyvinylpyrrolidone (as 10% solution in water) 4 mg Sodiumcarboxymethyl starch 4.5 mg Magnesium stearate 0.5 mg Talc 1 mg Total150 mg

The active ingredient, starch, and cellulose are passed through a No. 45mesh U.S. sieve, and the mixed thoroughly. The aqueous solutioncontaining polyvinylpyrrolidone is mixed with the resultant powder, andthe admixture then is passed through a No. 14 mesh U.S. sieve. Thegranules so produced are dried at 50° C. and passed through a No. 18mesh U.S. sieve. The sodium carboxymethyl starch, magnesium stearate,and talc, previously passed through No. 60 mesh U.S. sieve, are thenadded to the granules which, after mixing, are compressed on a tabletmachine to yield tablets each weighing 150 mg.

Formulation Example 5

Capsules, each containing 80 mg of active ingredient, are made asfollows:

Active ingredient 80 mg Starch 59 mg Microcrystalline cellulose 59 mgMagnesium stearate 2 mg Total 200 mg

The active ingredient, cellulose, starch, and magnesium stearate areblended, passed through a No. 45 mesh U.S. sieve, and filled into hardgelatin capsules in 200 mg quantities.

Formulation Example 6

Suppository, each containing 225 mg of active ingredient, are made asfollows:

Active ingredient  225 mg Saturated fatty acid glycerides 2000 mg Total2225 mg

The active ingredient is passed through a No. 60 mesh U.S. sieve andsuspended in the saturated fatty acid glycerides previously melted usingthe minimum heat necessary. The mixture is then poured into asuppository mold of nominal 2 g capacity and allowed to cool.

Formulation Example 7

Suspensions, each containing 50 mg of active ingredient per 5 ml dose,are made as follows:

Active ingredient 50 mg Sodium carboxymethyl cellulose 50 mg Syrup 1.25ml Benzoic acid solution 0.10 ml Flavor q.v. Color q.v. Purified waterto total 5 ml

The active ingredient is passed through a No. 45 U.S. sieve, and mixedwith the sodium carboxymethyl cellulose and syrup to form a smoothpaste. The benzoic acid solution, flavor and color are diluted with aportion of the water and added, with stirring. Sufficient water is thenadded to produce the required volume.

Formulation Example 8

An intravenous formulation may be prepared as follows:

Active ingredient  100 mg Isotonic saline 1000 ml

The solution of the above ingredients generally is administeredintravenously to a subject at a rate of 1 ml per minute.

Formulation Example 9

Composition of lyophilized preparations (in 1 vial) is made as follows:

Active ingredient 127 mg Trisodium citrate dihydrate  36 mg Mannitol 180mg

The above materials are dissolved in water for injection such that theconcentration of Active ingredient is 10 mg/g. The primary freezing stepis done for 3 hours at −40° C., the heat treating step for 10 hours at−10° C., and the re-freezing step for 3 hours at −40° C. Then, theprimary drying step is performed for 60 hours at 0° C., 10 Pa and thesecondary drying step for 5 hours at 60° C., 4 Pa. Thus the lyophilizedpreparation is obtained.

INDUSTRIAL APPLICABILITY

The compounds according to the present invention have sPLA₂ inhibitingactivity, so that the compounds of the invention inhibits sPLA₂-mediatedfatty acid (such as arachidonic acid) release, whereby it is effectivefor treating septic shock and the like.

What is claimed is:
 1. A compound represented by the formula (I):

wherein R¹ is hydrogen atom or a group selected from (a) C6 to C20alkyl, C6 to C20 alkenyl, C6 to C20 alkynyl, carbocyclic groups, andheterocyclic groups, (b) the groups represented by (a) each substitutedindependently with at least one group selected from non-interferingsubstituents, and (c) -(L¹)-R⁶ wherein L¹ is a divalent linking group of1 to 18 atom(s) selected from hydrogen atom(s), nitrogen atom(s), carbonatom(s), oxygen atom(s), and sulfur atom(s), and R⁶ is a group selectedfrom the groups (a) and (b); R² is hydrogen atom, or a group containing1 to 4 non-hydrogen atoms; R³ is -(L²)-(acidic group) wherein L² is anacid linker having an acid linker length of 1 to 5; R⁴ and R⁵ areselected independently from hydrogen atom, non-interfering substituents,carbocyclic groups, carbocyclic groups substituted with anon-interfering substituent(s), heterocyclic groups, and heterocyclicgroups substituted by a non-interfering substituent(s); and R^(A) is agroup represented by the formula:

wherein L⁷ is a divalent linker group selected from a bond or a divalentgroup selected from —CH₂—, —O—, —S—, —NH—, or —CO—, R²⁷ and R²⁸ areindependently hydrogen atom, C1 to C3 alkyl or a halogen; X and Y areindependently an oxygen atom or a sulfur atom; and Z is —NH₂ or —NHNH₂;a prodrug thereof, or its pharmaceutically acceptable salt, or itssolvate.
 2. A compound represented by the formula (II):

wherein R⁷ is hydrogen atom or —(CH₂)m—R¹² wherein m is an integer from1 to 6, and R¹² is (d) a group represented by the formula:

wherein a, c, e, n, q, and t are independently an integer from 0 to 2,R¹³ and R¹⁴ are independently selected from a halogen, C1 to C10 alkyl,C1 to C10 alkyloxy, C1 to C10 alkylthio, aryl, heteroaryl, and C1 to C10haloalkyl, α is an oxygen atom or a sulfur atom, L⁵ is —(CH₂)v—, —C═C—,—C≡C—, —O—, or —S—, v is an integer from 0 to 2, β is —CH₂— or —(CH₂)₂—,γ is an oxygen atom or a sulfur atom, b is an integer from 0 to 3, d isan integer from 0 to 4, f, p, and w are independently an integer from 0to 5, g is an integer from 0 to 2, r is an integer from 0 to 7, and u isan integer from 0 to 4, or is (e) a member of (d) substituted with atleast one substituent selected from the group consisting of C1 to C6alkyl, C1 to C6 alkyloxy, C1 to C6 haloalkyloxy, C1 to C6 haloalkyl,aryl, and a halogen; R⁸ is C1 to C3 alkyl, C1 to C3 alkenyl, C3 to C4cycloalkyl, C3 to C4 cycloalkenyl, C1 to C2 haloalkyl, C1 to C3alkyloxy, or C1 to C3 alkylthio; R⁹ is —(L³)—R¹⁵ wherein L³ isrepresented by the formula:

wherein M is —CH₂—, —O—, —N(R²⁴)—, or —S—, R¹⁶ and R¹⁷ are independentlyhydrogen atom, C1 to C10 alkyl, aryl, aralkyl, alkyloxy, haloalkyl,carboxy, or a halogen, and R²⁴ is hydrogen atom or C1 to C6 alkyl, andR¹⁵ is represented by the formula:

wherein R¹⁸ is hydrogen atom, a metal, or C1 to C10 alkyl, R19 isindependently hydrogen atom, or C1 to C10 alkyl, and t is an integerfrom 1 to 8; R¹⁰ and R¹¹ are independently hydrogen atom or anon-interfering substituent selected from hydrogen, C1 to C8 alkyl, C2to C8 alkenyl, C2 to C8 alkynyl, C7 to C12 aralkyl, C7 to C12 alkaryl,C3 to C8 cycloalkyl, C3 to C8 cycloalkenyl, phenyl, tolyl, xylyl,biphenyl, C1 to C8 alkyloxy, C2 to C8 alkenyloxy, C2 to C8 alkynyloxy,C2 to C12 alkyloxyalkyl, C2 to C12 alkyloxyalkyloxy, C2 to C12alkylcarbonyl, C2 to C12 alkylcarbonylamino, C2 to C12 alkyloxyamino, C2to C 12 alkyloxyaminocarbonyl, C1 to C12 alkylamino, C1 to C6 alkylthio,C2 to C12 alkylthiocarbonyl, C1 to C8 alkylsulfinyl, C1 to C8alkylsulfonyl, C2 to C8 haloalkyloxy, C1 to C8 haloalkylsulfonyl, C2 toC8 haloalkyl, C1 to C8 hydroxyalkyl, —C(O)O(C1 to C8 alkyl),—(CH₂)_(z)—O—(C1 to C8 alkyl), benzyloxy, aryloxy, arylthio,—(CONHSO₂R²⁵), —CHO, amino, amidino, halogen, carbamyl, carboxyl,carbalkoxy, —(CH₂)_(z)—CO₂H, cyano, cyanoguanidinyl, guanidino,hydrazide, hydrazino, hydrazido, hydroxy, hydroxyamino, iodo, nitro,phosphono, —SO₃H, thioacetal, thiocarbonyl, or carbonyl, R²⁵ is C1 to C6alkyl or aryl, z is an integer from 1 to 8; and R^(B) is a grouprepresented by the formula:

wherein Z is the same as defined above a prodrug thereof, or itspharmaceutically acceptable salt, or its solvate.
 3. A compound, aprodrug thereof, or its pharmaceutically acceptable salt, or its solvateas claimed in claim 1, wherein said R¹ is represented by the formula:

wherein R¹³, R¹⁴, b, d, f, g, p, r, u, w, α, β, and γ are the same asdefined above, L⁶ is a bond, —CH₂—, —C═C—, —C≡C—, —O—, or —S—.
 4. Acompound, a prodrug thereof, or its pharmaceutically acceptable salt, orits solvate as claimed in claim 1, wherein R² is C1 to C3 alkyl or C3 toC4 cycloalkyl.
 5. A compound, a prodrug thereof, or its pharmaceuticallyacceptable salt, or its solvate as claimed in claim 1, wherein L² is—O—CH₂—.
 6. A compound represented by the formula (III):

wherein R²⁰ is a group represented by the formula:

wherein L⁶, R¹³, R¹⁴, b, d, f, g, p, r, u, w, α, β, and γ are the sameas defined above, R²¹ is C1 to C3 alkyl or C3 to C4 cycloalkyl; L⁴ is—O—CH₂—, —S—CH₂—, —N(R²⁴)—CH₂—, —CH₂—CH₂—, —O—CH(CH₃)—, or—O—CH((CH₂)₂Ph)— wherein R²⁴ is hydrogen atom or C1 to C6 alkyl and Phis phenyl; R²² is —COOH, —SO₃H, or P(O)(OH)₂; R²³ is hydrogen atom, C1to C6 alkyl, C7 to C12 aralkyl, C1 to C6 alkyloxy, C1 to C6 alkylthio,C1 to C6 hydroxyalkyl, C2 to C6 haloalkyloxy, halogen, carboxy, C1 to C6alkyloxycarbonyl, aryloxy, arylthio, a carbocyclic group, or aheterocyclic group; and R^(B) is the same as defined above; a prodrugthereof, or its pharmaceutically acceptable salt, or its solvate.
 7. Acompound represented by the formula (IV):

wherein R²⁰, R²¹, R²³, and R^(B) are the same as defined above; and k isan integer from 1 to 3; a prodrug thereof, or its pharmaceuticallyacceptable salt, or its solvate.
 8. A compound, a prodrug thereof, orits pharmaceutically acceptable salt, or its solvate as claimed in claim6, wherein L⁴ is —O—CH₂—.
 9. A compound, a prodrug thereof, or itspharmaceutically acceptable salt, or its solvate as claimed in claim 1,wherein said R^(A) and R^(B) are -COCONH₂.
 10. A compound, a prodrugthereof, or its pharmaceutically acceptable salt, or its solvate asclaimed in claim 1, wherein R^(A) and R^(B) are —CH₂CONH₂.
 11. Acompound, a prodrug thereof, or its pharmaceutically acceptable salt, orits solvate as claimed in claim 1, wherein R^(A) and R^(B) are—CH₂CONHNH₂.
 12. The prodrug as claimed in claim 1 which is in the formof an ester.
 13. A pyrrolo[1,2-a]pyrazine compound selected from thegroup consisting of:[6-Benzyl-7-ethyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyacetic acid,[6-Cyclohexylmethyl-7-ethyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyaceticacid,[7-Ethyl-6-(3-methoxybenzyl)-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyaceticacid,[6-(Benzo[b]thiophen-6-ylmethyl)-7-ethyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyaceticacid,[6-Benzyl-7-ethyl-3-methyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyaceticacid,[7-Ethyl-6-(4-fluorobenzyl)-3-methyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyaceticacid, [6-(2-Biphenylmethyl)-7-ethyl-3-methyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyaceticacid,[6-Cyclopentylmethyl-7-ethyl-3-methyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyaceticacid, [6-(2-Benzyl)benzyl-7-ethyl-3-methyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyaceticacid,[7-Ethyl-6-(2-(4-fluorophenyl)benzyl)-3-methyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyaceticacid,[7-Ethyl-6-(3-fluorobenzyl)-3-methyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyaceticacid,[6-Benzyl-7-ethyl-3-isopropyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyaceticacid,[6-Benzyl-3,7-diethyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyaceticacid,[6-Benzyl-7-ethyl-8-oxamoyl-3-phenylpyrrolo[1,2-a]pyrazin-1-yl]oxyaceticacid,[6-Benzyl-7-ethyl-3-isobutyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyaceticacid,[3,6-Dibenzyl-7-ethyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyaceticacid,[7-Ethyl-3-methyl-8-oxamoyl-6-(2-(2-thienyl)benzyl)pyrrolo[1,2-a]pyrazin-1-yl]oxyaceticacid, [7-Ethyl-3-methyl-8-oxamoyl-6-(2-phenylethynylbenzyl)pyrrolo[1,2-a]pyrazin-1-yl]oxyacetic acid,[7-Ethyl-3-methyl-8-oxamoyl-6-( 2-phenyloxybenzyl)pyrrolo[1,2-a]pyrazin-1-yl]oxyacetic acid,[7-Ethyl-3-methyl-8-oxamoyl-6-(2-(3-thienyl)benzyl)pyrrolo[1,2-a]pyrazin-1-yl]oxyacetic acid,[7-Ethyl-3-methyl-6-(2-(5-methylthien-2-yl)benzyl)-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyaceticacid,[7-Ethyl-6-(2-(4-methoxyphenyl)benzyl)-3-methyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyaceticacid,[7-Ethyl-3-methyl-6-(2-(4-methylphenyl)benzyl)-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyaceticacid, [7-Ethyl-3-methyl-8-oxamoyl-6-(2-(2-phenylethyl)benzyl)pyrrolo[1,2-a[pyrazin-1-yl]oxyacetic acid,[6-Benzyl-7-cyclopropyl-3-methyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyaceticacid,[7-Cyclopropyl-6-(4-fluorobenzyl)-3-methyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyaceticacid,[6-Benzyl-3-cyclohexyl-7-ethyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyaceticacid,[6-(2-Biphenylmethyl)-3-cyclohexyl-7-ethyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyaceticacid,[6-Benzyl-3,7-dimethyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyaceticacid,[7-Ethyl-3-methyl-6-(5-methylthien-2-ylmethyl)-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyaceticacid,[6-(Benzo[b]thiophen-3-ylmethyl)-7-ethyl-3-methyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyaceticacid, Sodium[7-ethyl-6-(4-fluorobenzyl)-3-methyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyacetate,Sodium[7-ethyl-6-(2-(4-fluorophenyl)benzyl)-3-methyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyacetate,Sodium [7-ethyl-3-methyl-8-oxamoyl-6-(2-(2-thienyl) benzyl)pyrrolo[1,2-a]pyrazin-1-yl]oxyacetate, Sodium[7-ethyl-3-methyl-8-oxamoyl-6-(2-(3-thienyl)benzyl)pyrrolo[1,2-a]pyrazin-1-yl]oxyacetate, and a prodrug thereof, orits pharmaceutically acceptable salt, or its solvate.
 14. Apyrrolo[1,2-a] pyrazine compound selected from the group consisting ofMethyl [7-ethyl-6-(2-(4-fluorophenyl)benzyl)-3-methyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1 -yl]oxyacetate, Ethyl[7-ethyl-6-(2-(4-fluorophenyl)benzyl)-3-methyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyacetate, Morpholinylethyl[7-ethyl-6-(2-(4-fluorophenyl)benzyl)-3-methyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyacetate,Sodium[7-ethyl-6-(2-(4-fluorophenyl)benzyl)-3-methyl-8-oxamoylpyrrolo[1,2-a]pyrazin-1-yl]oxyacetate, Methyl[7-ethyl-3-methyl-8-oxamoyl-6-(2-(2-thienyl)benzyl)pyrrolo[1,2-a]pyrazin-1-yl]oxyacetate,Ethyl[7-ethyl-3-methyl-8-oxamoyl-6-(2-(2-thienyl)benzyl)pyrrolo[1,2-a]pyrazin-1-yl]oxyacetate,Morpholinylethyl[7-ethyl-3-methyl-8-oxamoyl-6-(2-(2-thienyl)benzyl)pyrrolo(1,2-a]pyrazin-1-yl]oxyacetate, and Sodium[7-ethyl-3-methyl-8-oxamoyl-6-(2-(2-thienyl)benzyl)pyrrolo[1,2-a]pyrazin-1-yl]oxyacetate.15. A pharmaceutical composition containing a compound as claimed inclaim 1 as an active ingredient.
 16. A pharmaceutical composition asclaimed in claim 15, wherein said composition is for inhibiting sPLA₂.17. A pharmaceutical composition as claimed in claim 15, wherein saidcomposition is for treatment or prevention of Inflammatory Diseases. 18.A method of inhibiting sPLA₂ mediated release of fatty acid whichcomprises contacting sPLA₂ with a therapeutically effective amount of apyrrolo[1,2-a]pyrazine compound as claimed in claim
 1. 19. A method oftreating a mammal to alleviate the pathological effects of InflammatoryDiseases; wherein the method comprises administration to said mammal ofa pyrrolo[1,2-a]pyrazine compound as claimed in claim 1 in apharmaceutically effective amount.
 20. A compound of claim 1 or apharmaceutical formulation containing an effective amount of apyrrolo[1,2-a]pyrazine compound of claim 1 for use in treatment ofInflammatory Diseases.
 21. A compound of claim 1 or a pharmaceuticalformulation containing an effective amount of a pyrrolo[1,2-a]pyrazinecompound of claim 1 for use as an inhibitor for inhibiting sPLA₂mediated release of fatty acid.
 22. A compound, a prodrug thereof, orits pharmaceutically acceptable salt or its solvate as claimed in claim2, wherein said R⁷is represented by the formula:

wherein R¹³, R¹⁴, b, d, f, g, p, r, u, w, α, β, and γ are the same asdefined in claim 2, L⁶ is a bond, —CH₂—, —C═C—, —C≡C—, —O—, or —S—. 23.A compound, a prodrug thereof, or its pharmaceutically acceptable salt,or its solvate as claimed in claim 2, wherein R⁸ is C1 to C3 alkyl or C3to C4 cycloalkyl.
 24. A compound, a prodrug thereof, or itspharmaceutically acceptable salt, or its solvate as claimed in claim 2,wherein L³is —O—CH₂—.